Name: Anti-GAPDH antibody [EPR16891] - Loading Control
SKU: ab181602
Rating: 5 (23 reviews)

Anti-GAPDH antibody [EPR16891] - Loading Control is a rabbit recombinant monoclonal antibody that is used to detect GAPDH in Flow cytometry (Intra), ICC/IF, IHC-P, IP, Western blot. Suitable for African green monkey, Chicken, Human, Mouse, Rat, Xenopus tropicalis, Zebrafish samples.

- Recombinant format for unrivaled batch-batch consistency
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Cited in over 2400 publications

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Tested	Expected	Expected	Tested
Rat	Expected	Tested	Expected	Expected	Tested
African green monkey	Expected	Tested	Expected	Expected	Expected
Chicken	Expected	Tested	Expected	Expected	Expected
Fish	Predicted	Predicted	Predicted	Predicted	Predicted
Rabbit	Predicted	Predicted	Predicted	Predicted	Predicted
Xenopus tropicalis	Expected	Tested	Expected	Expected	Expected
Zebrafish	Expected	Tested	Expected	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/60	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat, African green monkey, Zebrafish, Xenopus tropicalis, Chicken	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rabbit, Fish	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/10000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes -
Species African green monkey	Dilution info 1/10000	Notes -
Species Zebrafish	Dilution info 1/10000	Notes -
Species Xenopus tropicalis	Dilution info 1/10000	Notes -
Species Chicken	Dilution info 1/10000	Notes -
Species Human	Dilution info 1/10000	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rabbit, Fish	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat, African green monkey, Zebrafish, Xenopus tropicalis, Chicken	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rabbit, Fish	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/180	Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat, African green monkey, Zebrafish, Xenopus tropicalis, Chicken	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rabbit, Fish	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species African green monkey, Zebrafish, Xenopus tropicalis, Chicken	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rabbit, Fish	Dilution info -	Notes -

Associated Products

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Target data

See full target information GAPDH

Function

Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively (PubMed:11724794, PubMed:3170585). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate (PubMed:11724794, PubMed:3170585). Modulates the organization and assembly of the cytoskeleton (By similarity). Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes (PubMed:23071094). Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation (PubMed:23071094). Also plays a role in innate immunity by promoting TNF-induced NF-kappa-B activation and type I interferon production, via interaction with TRAF2 and TRAF3, respectively (PubMed:23332158, PubMed:27387501). Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis (By similarity). Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity).

Alternative names

GAPD, CDABP0047, OK/SW-cl.12, GAPDH, Glyceraldehyde-3-phosphate dehydrogenase, Peptidyl-cysteine S-nitrosylase GAPDH

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR16891
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P, IP and WB.

Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) was first used in a scientific publication in 1987 and has been cited over 2472 times in peer reviewed journals. It's performance in Western Blot in human, mouse and rat samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) has 22 independent reviews from customers.

GAPDH antibodies are often used as loading controls in Western Blot. Anti-GAPDH antibody [EPR16891] - Loading Control has been verified in Western Blot samples and detects a band at 36kDa Molecular weight.

Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) specifically detects GAPDH (UniProt ID: P16858; Molecular weight: 36kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPR16891 - Anti-GAPDH antibody [EPR16891] - BSA and Azide free ab199553.

Antibody clone EPR16891 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 647, Alexa Fluor^® 488, HRP, Alexa Fluor^® 594, Alexa Fluor^® 405, PE, FITC (Alexa Fluor® 647 Anti-GAPDH antibody [EPR16891] ab201272, Alexa Fluor® 488 Anti-GAPDH antibody [EPR16891] ab201768, HRP Anti-GAPDH antibody [EPR16891] - Loading Control ab201822, Alexa Fluor® 594 Anti-GAPDH antibody [EPR16891] ab206371, Alexa Fluor® 405 Anti-GAPDH antibody [EPR16891] ab206372, PE Anti-GAPDH antibody [EPR16891] ab224004, FITC Anti-GAPDH antibody [EPR16891] ab224005).

One of the top cited clones in the market for GAPDH with >3000 citations. GAPDH is a key target involved in glycolysis and cell metabolism. It plays a crucial role as a housekeeping gene, particularly in understanding GAPDH expression and its function as a metabolic enzyme. GAPDH is widely analysed in GAPDH activity assays and studies of its role in various cellular processes. Additionally, GAPDH is commonly used as a loading control in western blotting, IHC and immunofluorescence experiments.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Glyceraldehyde-3-phosphate dehydrogenase commonly known as GAPDH is an enzyme involved in glycolysis. Its molecular weight (MW) is approximately 36 kDa. The protein is expressed ubiquitously in almost all tissues reflecting its essential role in energy production. GAPDH catalyzes the sixth step of glycolysis converting glyceraldehyde-3-phosphate into 13-bisphosphoglycerate. Due to its stable expression researchers often use GAPDH as a loading control in western blot experiments.

Biological function summary

GAPDH serves important metabolic functions beyond its enzymatic role in glycolysis. It functions as part of a multi-enzyme complex within the cytoplasm which facilitates efficient substrate channeling during glycolysis. Additionally GAPDH has non-glycolytic roles including involvement in nuclear processes like RNA export and DNA repair. Its ubiquitous presence across different cellular compartments indicates its multiple functions beyond metabolic pathways.

Pathways

GAPDH integrates into significant cellular functions like the glycolytic pathway and apoptotic pathways. In glycolysis GAPDH collaborates with enzymes like phosphoglycerate kinase forming a cohesive link in the energy conversion chain. Its participation in apoptotic pathways highlights GAPDH's involvement in cellular death processes interacting with proteins like Bcl-2 to influence apoptosis progression. These roles reinforce its presence in central metabolic and regulatory pathways.

Associated diseases and disorders

GAPDH has associations with neurodegenerative diseases and cancer. In neurodegenerative disorders such as Alzheimer's disease GAPDH’s altered enzymatic activity is frequently observed influencing cellular energy homeostasis. Moreover overexpression or aberrant regulation of GAPDH relates to cancer cell proliferation and metastasis implicating proteins like p53 in these pathways. The diverse functions and interactions of GAPDH emphasize its importance in both normal cellular function and disease states.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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41 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181602 at 1/60 dilution. Western blot was performed from the immunoprecipitate using ab181602 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract.
Lane 2: PBS instead of HeLa whole cell extract.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Predicted band size: 36 kDa
Observed band size: 36 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Lanes 1-3: Merged signal (red and green). Green - ab88960 observed at 27 kDa. Red - loading control ab181602 observed at 36 kDa.
ab88960 Anti-Cathepsin H antibody [AF14D7] was shown to specifically react with Cathepsin H in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human CTSH (Cathepsin H) knockout HEK-293T cell line ab266245 (knockout cell lysate Human CTSH (Cathepsin H) knockout HEK-293T cell lysate ab258387) was used. Wild-type and Cathepsin H knockout samples were subjected to SDS-PAGE. ab88960 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Anti-Cathepsin H antibody [AF14D7] (ab88960) at 1/1000 dilution
Lane 1: Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: CTSH knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 3: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Secondary
Lanes 1 - 3: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Lanes 1 - 3: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution
Observed band size: 37 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution
Lane 1: Mouse kidney lysates at 10 µg
Lane 2: Mouse spleen lysates at 10 µg
Lane 3: RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates at 10 µg
Lane 4: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 10 µg
Lane 5: Rat brain lysates at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells labeling GAPDH with ab181602 at dilution of 1/500. The secondary antibody used was Alexa Fluor^® 488; goat anti-rabbit IgG (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400. Nucleus was counter-stained with DAPI (blue). Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor^®594 goat anti-mouse secondary, 1/500). The negative controls are shown in the bottom middle and right hand panels- for negative control 1 primary antibody (ab181602; 1/500) and secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120; 1/500) was used. For negative control 2 primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291; 1/500) and secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077; 1/400) was used.
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
All lanes: Western blot - Anti-METTL3 antibody [EPR18810] (Anti-METTL3 antibody [EPR18810] ab195352) at 1/1000 dilution
Lane 1: Mouse brain lysate prepared in 1%SDS Hot lysis method at 20 µg
Lane 2: Mouse brain lysate prepared in RIPA lysis method at 20 mg/mL
Lane 3: Mouse heart lysate prepared in 1%SDS Hot lysis method at 20 µg
Lane 4: Mouse heart lysate prepared in RIPA lysis method at 20 mg/mL
Lane 5: Mouse kidney lysate prepared in RIPA lysis method at 20 µg
Lanes 6 - 7: Mouse spleen lysate prepared in spleen lysis method at 20 µg
Lane 8: Rat brain lysate prepared in RIPA lysis method at 20 µg
Lane 9: Rat kidney lysate prepared in RIPA lysis method at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 64 kDa
Observed band size: 64 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
False colour image of Western blot: Anti-BRAF (mutated V600E) antibody [VE1] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-BRAF (mutated V600E) antibody [VE1] ab228461 was shown to bind specifically to mutant BRAF V600E. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed at 1/20000 dilution.
This image was generated in-house using a previous batch, manufactured using hybridoma production method.
All lanes: Western blot - Anti-BRAF (mutated V600E) antibody [VE1] (Anti-BRAF (mutated V600E) antibody [VE1] ab228461) at 1/1000 dilution
Lane 1: HCT 116 cell lysate (wildtype BRAF) at 20 µg
Lane 2: SW480 cell lysate (wildtype BRAF) at 20 µg
Lane 3: Caco-2 cell lysate (wildtype BRAF) at 20 µg
Lane 4: HT-29 cell lysate (mutant BRAF V600E) at 20 µg
Lane 5: A375 cell lysate (mutant BRAF V600E) at 20 µg
Secondary
Lanes 1 - 5: Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed at 1/20000 dilution
Lanes 1 - 5: Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 84 kDa
Observed band size: 85 kDa, 36 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
All lanes: Western blot - Anti-ErbB2 / HER2 antibody [SP101] (Anti-ErbB2 / HER2 antibody [SP101] ab231438)
All lanes: SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 0.05 µg/mL
Predicted band size: 137 kDa
Observed band size: 180 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour before being incubated with Anti-GAPDH antibody ab181602 overnight at 4°C. Antibody binding was detected using the Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
Image shows merged signal (red and green). Green - Anti-beta Actin antibody [AC-15] (Anti-beta Actin antibody [AC-15] - Loading Control ab6276)Anti-beta Actin antibody [AC-15] - Loading Control ab6276 observed at 42 kDa. Red - GAPDH loading control, ab181602, observed at 37 kDa.
Lanes 1 - 2: Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (Anti-beta Actin antibody [AC-15] - Loading Control ab6276) at 1/5000 dilution
Lanes 1 - 2: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Beta actin knockout HAP1 cell lysate (20 µg)
Secondary
Lanes 1 - 2: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/10000 dilution
Lanes 1 - 2: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Predicted band size: 36 kDa, 41 kDa
Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181602 at 1/180 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/50000 dilution
Lane 1: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 20 µg
Lane 2: Xenopus tropicalis muscle lysates at 20 µg
Lane 3: UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates at 20 µg
Lane 4: Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Different batches of ab181602 were tested on HeLa (Human cervix adenocarcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 36 kDa.
All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Predicted band size: 36 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of human bladder is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution
Lane 1: COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates at 20 µg
Lane 2: Zebrafish lysates at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution
Lane 1: Human fetal brain lysates at 10 µg
Lane 2: Human fetal heart lysates at 10 µg
Secondary
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 15964844).
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-NOR1/TEC antibody [EPR26986-37] (Anti-NOR1/TEC antibody [EPR26986-37] ab313781) at 1/1000 dilution
Lane 1: Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg
Lane 2: THP-1 treated first with 80nM TPA for 24 hours, then replaced with /ml LPS for 6 hours whole cell lysate at 20 µg
Lane 3: Hela (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 50 µg
Lane 4: Hela transfected with siRNA specifically targeti NOR1/TEC whole cell lysate at 50 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 75 kDa, 80 kDa
Exposure time: 180s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The 120, 140 and 180kDa bands are different isoforms as reported in the literature (PMID:26288071).
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-NCAM1 antibody [EPR26939-108] (Anti-NCAM1 antibody [EPR26939-108] ab313779) at 1/1000 dilution
Lane 1: Human hippocampus tissue lysate at 20 µg
Lane 2: Human cerebellum tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 120 kDa, 140 kDa, 180 kDa
Exposure time: 15s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 30102396, 28783698, 12368356).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-SP140 antibody [EPR25120-83] (Anti-SP140 antibody [EPR25120-83] ab314211) at 1/1000 dilution
Lane 1: HuT-78 (human Sezary syndrome cutaneous T lymphocyte) transfected with scrambled siRNA control whole cell lysate at 20 µg
Lane 2: HuT-78 transfected with siRNA specifically targeti SP140 whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 140 kDa, 130 kDa
Exposure time: 180s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: MCF7 (PMID: 18772882).
The 120, 140 and 180kDa bands are different isoforms as reported in the literature (PMID:26288071)
The identity of the lower MW bands at approximately 75 kDa and 50kDa are unknown.
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
Exposure time: Lane 1: 147 seconds; Lane 2, 3: 92 seconds.
All lanes: Western blot - Anti-NCAM1 antibody [EPR26939-108] (Anti-NCAM1 antibody [EPR26939-108] ab313779) at 1/1000 dilution
Lane 1: No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) whole cell lysate at 20 µg
Lane 2: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 120 kDa, 140 kDa, 180 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-SP140 antibody [EPR25120-83] (Anti-SP140 antibody [EPR25120-83] ab314211) at 1/1000 dilution
Lane 1: HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate at 20 µg
Lane 2: Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg
Lane 3: Ramos (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under non-reducing conditions.
Observed band size: 140 kDa, 130 kDa
Exposure time: 81s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: liver, testis(PMID: 8929530)
Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
Samples are non-boiled as boiling may cause protein aggregation.
The identity of the lower MW band at approximately 100 kDa may represent an N-terminal truncated isoform of CACNA1A (PMID: 7673157).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Pan Cav2 antibody [EPR28073-84] (Anti-Pan Cav2 antibody [EPR28073-84] ab315092) at 1/1000 dilution
Lane 1: Mouse brain tissue lysate at 20 µg
Lane 2: Mouse liver tissue lysate at 20 µg
Lane 3: Mouse testis tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under non-reducing conditions.
Observed band size: 100 kDa, 267 kDa, 36 kDa
Exposure time: 180s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 27696294).
Low expression: testis (PMID: 18455983).
The identity of the band at 10 kDa is unknown.
Lanes 5-8 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 and lanes 1-4 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-TIPE2 antibody [EPR27964-44] (Anti-TIPE2 antibody [EPR27964-44] ab324268) at 1/1000 dilution
Lane 1: Human lung cancer tissue lysate at 20 µg
Lane 2: Human lymph node tissue lysate at 20 µg
Lane 3: Human tonsil tissue lysate at 20 µg
Lane 4: Human testis tissue lysate at 20 µg
Lane 5: Mouse thymus tissue lysate at 20 µg
Lane 6: Mouse lymph node tissue lysate at 20 µg
Lane 7: Mouse spleen tissue lysate at 20 µg
Lane 8: Mouse testis tissue lysate at 20 µg
Secondary
Lanes 1 - 4: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Lanes 5 - 8: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 16 kDa, 18 kDa, 36 kDa
Exposure time: 26s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: A20, EL4.
To minimize protein degradation, the fresh lysates were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.
The identity of the bands higher than 75 kDa are unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Deptor antibody [EPR30295-507] (ab324271) at 1/1000 dilution
Lane 1: C2C12 (mouse myoblast) whole cell lysate at 30 µg
Lane 2: C2C12 (mouse myoblast) whole cell lysate (fresh lysates) at 30 µg
Lane 3: A20 (mouse reticulum sarcoma b lymphocyte) whole cell lysate (fresh lysates) at 30 µg
Lane 4: EL4 (mouse lymphoma t lymphocyte) whole cell lysate at 30 µg
Lane 5: F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 30 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/2000 dilution
Observed band size: 48 kDa, 34 kDa, 36 kDa
Exposure time: 15s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: colon, stomach.
The molecular weight observed is consistent with what has been described in the literature (PMID: 29285063, 19726871).
Samples are non-boiled as boiling may cause protein aggregation.
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
Exposure time: Lane 1-3: 37 seconds; Lane 4: 10 seconds.
All lanes: Western blot - Anti-Cx40/GJA5 antibody [EPR28370-62] (Anti-Cx40/GJA5 antibody [EPR28370-62] ab313644) at 1/1000 dilution
Lane 1: Mouse heart tissue lysate at 20 µg
Lane 2: Mouse colon tissue lysate at 20 µg
Lane 3: Mouse stomach tissue lysate at 20 µg
Lane 4: Mouse lung tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under non-reducing conditions.
Observed band size: 40 kDa, 36 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
SLC12A1/NKCC2 is a glycoprotein of approximately 150 kDa and detected as a 135-kDa band after treated with deglycosylation treatment.
The expression profile/ molecular weight observed is consistent with what has been described in literature (PMID: 22759959).
Samples are non-boiled as boiling may cause protein aggregation.
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-SLC12A1/NKCC2 antibody [EPR28176-90] (Anti-SLC12A1/NKCC2 antibody [EPR28176-90] ab313640) at 1/1000 dilution
Lane 1: Untreated Mouse kidney tissue lysate at 20 µg
Lane 2: Mouse kidney tissue lysate treated with Peptide:N-glycosidase F (PNGase F) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under non-reducing conditions.
Observed band size: 135 kDa, 150 kDa, 270 kDa
Exposure time: 15s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 29285063, 19726871).
Samples are non-boiled as boiling may cause protein aggregation.
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Anti-Integrin alpha V (Anti-Integrin alpha V antibody [EPR16800] ab179475) control staining at 1/5000 dilution.
All lanes: Western blot - Anti-Cx40/GJA5 antibody [EPR28370-62] (Anti-Cx40/GJA5 antibody [EPR28370-62] ab313644) at 1/1000 dilution
Lane 1: Mouse lung non-membrane fraction at 20 µg
Lane 2: Mouse lung membrane fraction at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under non-reducing conditions.
Observed band size: 40 kDa, 36 kDa
Exposure time: 37s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: Raji, HDLM-2 (PMID: 26066800).
The expression molecular weight observed is consistent with what has been described in the literature (PMID: 14673177).
In lanes 1-3, the lysates were stored at -80°C prior to Western Blotting. The bands beneath the target band (140 kDa) are likey to be degradation products. In lanes 4-5, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-CSF-1-R antibody [EPR28407-22] (Anti-CSF-1-R antibody [EPR28407-22] ab313648) at 1/1000 dilution
Lanes 1 and 4: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg
Lanes 2 and 5: Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg
Lane 3: HDLM-2 (human Hodgkin lymphoma) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 190 kDa, 140 kDa
Exposure time: 180s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 27002480).
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-GM2A antibody [EPR28371-75] (Anti-GM2A antibody [EPR28371-75] ab313587) at 1/1000 dilution
Lane 1: SK-BR-3 (human breast adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg
Lane 2: SK-BR-3 transfected with siRNA specifically targeti GM2A whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 20 kDa, 21 kDa
Exposure time: 15s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: MCF7 (PMID: 27002480).
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 27002480).
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
All lanes: Western blot - Anti-GM2A antibody [EPR28371-75] (Anti-GM2A antibody [EPR28371-75] ab313587) at 1/1000 dilution
Lane 1: SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 20 kDa, 21 kDa
Exposure time: 26s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: 293T (PMID: 34880314).
All lanes: Western blot - Anti-EZHIP antibody [EPR28396-50] (Anti-EZHIP antibody [EPR28396-50] ab313392) at 1/1000 dilution
Lane 1: U-2OS (human bone osteosarcoma epithelial cell) whole cell lysate at 50 µg
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 50 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under non-reducing conditions.
Observed band size: 50, kDa, 75 kDa
Exposure time: 92s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: spleen (PMID: 36535626).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Deptor antibody [EPR30295-507] (ab324271) at 1/1000 dilution
Lane 1: Mouse pancreas tissue lysate at 20 µg
Lane 2: Mouse spleen tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 48 kDa, 34 kDa, 36 kDa
Exposure time: 15s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.
Exposure time: Lanes 1-2: 26 seconds; Lane 3: 136 seconds
All lanes: Western blot - Anti-Deptor antibody [EPR30295-507] (ab324271) at 1/1000 dilution
Lane 1: Mouse lung tissue lysate at 20 µg
Lane 2: Mouse skeletai muscle tissue lysate at 20 µg
Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 48 kDa, 34 kDa, 36 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
TNFRSF14/HVEM is a glycoprotein of approximately 30 kDa and 50 kDa and detected as a 20 kDa and 40 kDa band after treated with Protein Deglycosylation MIX II.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-TNFRSF14/HVEM antibody [EPR28154-164] (Anti-TNFRSF14/HVEM antibody [EPR28154-164] ab314494) at 1/1000 dilution
Lane 1: Untreated SK-MEL-28 (human malignant melanoma) whole cell lysate at 50 µg
Lane 2: SK-MEL-28 treated with Protein Deglycosylation Mix II at 50 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 20 kDa, 30 kDa, 40 kDa, 50 kDa
Exposure time: 92s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.
The identity of the bands lower than 78 kDa are unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-DNA polymerase eta antibody [EPR29670-68] (ab324350) at 1/1000 dilution
Lane 1: HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 48 µg
Lane 2: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 48 µg
Lane 3: U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg
Lane 4: MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 78 kDa, 36 kDa
Exposure time: 180s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-MTHFD1 antibody [EPR29810-508] (ab324379) at 1/1000 dilution
Lane 1: HEK-293 (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg
Lane 2: HEK-293 transfected with siRNA specifically targeting MTHFD1 whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 101 kDa, 36 kDa
Exposure time: 15s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-Beta Arrestin 2 antibody [EPR25410-70] ab314213 was shown to bind specifically to Beta Arrestin 2. A band was observed at 50 kDa and 37kDa in wild-type 293T cell lysates whereas loss of signal was observed in the ARRB2 knockout cell line Human ARRB2 (Beta Arrestin 2) knockout HEK-293T cell line ab266116 (knockout cell lysate Human ARRB2 (Beta Arrestin 2) knockout HEK-293T cell lysate ab257282)./p>
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Beta Arrestin 2 antibody [EPR25410-70] (Anti-Beta Arrestin 2 antibody [EPR25410-70] ab314213) at 1/1000 dilution
Lane 1: Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 2: Western blot - Human ARRB2 (Beta Arrestin 2) knockout HEK-293T cell lysate (Human ARRB2 (Beta Arrestin 2) knockout HEK-293T cell lysate ab257282) at 20 µg
Lane 3: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 37,50 kDa
Exposure time: 180s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Western blot: Mouse Monoclonal [CL4438] to USP30 Anti-USP30 antibody [CL4438] ab219969 staining at 1/1000 dilution, shown in green; Rabbit anti GAPDH (ab181602) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 59 kDa in Wild-type A549 ab288558 cell lysates with no signal observed at this size in USP30 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.
All lanes: Western blot - Anti-USP30 antibody [CL4438] (Anti-USP30 antibody [CL4438] ab219969) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: Western blot - Human USP30 Knockout A549 cell line (Human USP30 Knockout A549 cell line ab324260) at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Secondary
All lanes: Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 59 kDa
Observed band size: 59 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: liver, lung, pancreas, spleen (PMID: 15883926).
The identity of the bands lower than 37 kDa are unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-KIAA1279 antibody [EPR30401-526] (Anti-KIAA1279 antibody [EPR30401-526] ab324162) at 1/1000 dilution
Lane 1: Mouse brain tissue lysate at 50 µg
Lane 2: Mouse lung tissue lysate at 50 µg
Lane 3: Mouse liver tissue lysate at 50 µg
Lane 4: Mouse pancreas tissue lysate at 50 µg
Lane 5: Mouse testis tissue lysate at 50 µg
Lane 6: Mouse spleen tissue lysate at 50 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 75 kDa, 36 kDa
Exposure time: 37s
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Negative control: Neuro-2a (PMID: 10779524)
Low expression: liver, skin (PMID: 10779524)
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 10779524).
The identity of the band at approximately 15 kDa in lane 1 is likely to be N-Term target fragments..
The identity of the band at approximately 100 kDa is unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
Exposure time: Lanes 1-2: 59 seconds; Lanes 3-5: 114 seconds
All lanes: Western blot - Anti-Dectin-1 antibody [EPR29712-507] (Anti-Dectin-1 antibody [EPR29712-507] ab324238) at 1/1000 dilution
Lane 1: N9 (mouse microglia cell) whole cell lysate at 40 µg
Lane 2: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 40 µg
Lane 3: Mouse spleen tissue lysate at 40 µg
Lane 4: Mouse liver tissue lysate at 40 µg
Lane 5: Mouse skin tissue lysate at 40 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 25-40 kDa, 36 kDa
Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
GAPDH Western blot staining using rabbit Anti-GAPDH antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
Exposure time: Lanes 1-4: 48 seconds, lanes 5-10: 180 seconds
All lanes: Western blot - Anti-Density Regulated Protein antibody [EPR29751-532] (Anti-Density Regulated Protein antibody [EPR29751-532] ab324269) at 1/1000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 2: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg
Lanes 3 - 4: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
Lane 5: MEF (mouse embryo fibroblast) whole cell lysate at 20 µg
Lane 6: P815 (mouse mastocytoma mast Cell) whole cell lysate at 20 µg
Lane 7: F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 8: EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg
Lane 9: Mouse heart tissue lysate at 20 µg
Lane 10: Mouse kidney tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 26 kDa, 36 kDa