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Anti-GAPDH antibody [EPR16891] ab181602 is a rabbit monoclonal antibody that is used in GAPDH western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR16891 is cited in over 3150 publications
- One antibody for all your GAPDH staining, use in GAPDH western blotting, IHC, immunofluorescence and flow cytometry

Same trusted quality, new lower price


Images

Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602), expandable thumbnail
  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602), expandable thumbnail
  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (AB181602), expandable thumbnail
  • 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
IPWBICC/IFFlow 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

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

Mouse, Rat, African green monkey, Zebrafish, Xenopus tropicalis, Chicken

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rabbit, Fish

Dilution info

-

Notes

-

Tested
Tested

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

Rabbit, Fish

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/500

Notes

-

Expected
Expected

Species

Mouse, Rat, African green monkey, Zebrafish, Xenopus tropicalis, Chicken

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rabbit, Fish

Dilution info

-

Notes

-

Tested
Tested

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

Mouse, Rat, African green monkey, Zebrafish, Xenopus tropicalis, Chicken

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rabbit, Fish

Dilution info

-

Notes

-

Tested
Tested

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

African green monkey, Zebrafish, Xenopus tropicalis, Chicken

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Rabbit, Fish

Dilution info

-

Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

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

Recommended products

Anti-GAPDH antibody [EPR16891] ab181602 is a rabbit monoclonal antibody that is used in GAPDH western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR16891 is cited in over 3150 publications
- One antibody for all your GAPDH staining, use in GAPDH western blotting, IHC, immunofluorescence and flow cytometry

Same trusted quality, new lower price

Key facts

Isotype

IgG

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
Loading...

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

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

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:
Terms & Conditions.

37 product images

  • Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    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.

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    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] ab6276)Anti-beta Actin antibody [AC-15] 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] (Anti-beta Actin antibody [AC-15] 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

    Lane 1: Wild-type HAP1 cell lysate (20 μg)
    Lane 2: Beta actin knockout HAP1 cell lysate (20 μg)

    Lanes 1 and 2: Merged signal (red and green). Green - beta actin, Anti-beta Actin antibody [AC-15] ab6276 observed at 42 kDa. Red - loading control, ab181602 observed at 37 kDa.

    Anti-beta Actin antibody [AC-15] ab6276 was shown to specifically react with beta actin in wild-type HAP1 cells. No band was observed when beta actin knockout samples were used. Wild-type and beta actin knockout samples were subjected to SDS-PAGE. Anti-beta Actin antibody [AC-15] ab6276 (beta actin) and ab181602 (loading control to GAPDH) were diluted 1/5000 and 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Low expression tissue: skeletal muscle

    The expression molecular weight observed is consistent with what has been described in the literature (PMID: 8373947).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

    Lane 1: Human testis tissue lysate at 20 µg

    Lane 2: Human tonsil tissue lysate at 20 µg

    Lane 3: Human skeletal muscle 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

    Exposure time: 3s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Negative control: Jurkat

    In Western blot, ab323182 was shown to bind specifically to Clusterin. Target of interest was observed at 62 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in Clusterin knockout cell line (lane 2).

    The expression molecular weight observed is consistent with what has been described in the literature (PMID: 8373947).

    Recombinant multiclonal ab323182 performs better than polyclonal in WB application.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-Clusterin antibody [RM1292] (ab323182) at 1/1000 dilution

    Lane 1: Wide-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: Clusterin knockout HeLa whole cell lysate at 20 µg

    Lane 3: Huh7 (human hepatocelullar carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Negative control: Jurkat.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 21673655, PMID: 24560577).

    The identity of the bands around 50 kDa and 37KDa in lane 3 are unkown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-FLRT3 antibody [EPR29132-87] (ab323410) at 1/1000 dilution

    Lane 1: A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: LN-229 (human brain glioblastoma epithelial cell) whole cell lysate at 20 µg

    Lane 3: Huh7 (human hepatocelullar carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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: 75-100 kDa, 36 kDa

    Exposure time: 114s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The identity of the lower MW band at approximately 15 kDa (in lane 6) is unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    Exposure time: Lanes 1-2: 37 seconds, lanes 3-6: 180 seconds

    All lanes: Western blot - Anti-FOXO3A antibody [EPR29854-587] (ab323467) at 1/1000 dilution

    Lane 1: Mouse liver tissue lysate at 20 µg

    Lane 2: Mouse testis tissue lysate at 20 µg

    Lane 3: Mouse pancreas tissue lysate at 20 µg

    Lane 4: Mouse heart tissue lysate at 20 µg

    Lane 5: Mouse brain tissue lysate at 20 µg

    Lane 6: Rat brain 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

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

    In Western blot, Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture) ab242509 was shown to bind specifically to MIF. Target of interest was observed at 13 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in MIF knockout cell line (lane 2).

    Exposure time: Lane 1-4: 180 seconds
    Lane 5-8: 15 seconds

    All lanes: Western blot - Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture) (Anti-MIF antibody [EPR18149-132] - BSA and Azide free (Capture) ab242509) at 1/1000 dilution

    Lane 1: Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

    Lane 2: MIF knockout HAP1 whole cell lysate at 20 µg

    Lane 3: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

    Lane 4: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 5: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

    Lane 6: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

    Lane 7: J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate at 20 µg

    Lane 8: RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) 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: 13 kDa

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    In Western blot, ab323371 was shown to bind specifically to TNFRSF10B. Target of interest was observed at 40, 48 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in TNFRSF10B knockout cell line (lane 2) (lane 2, knockout cell line Human TNFRSF10B (DR5) knockout HeLa cell line ab264922).

    The lanes 1-5 were developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

    Doxorubicin treatment elevated the expression of DR5 (PMID: 12496481; PMID: 11468181; PMID: 11090076).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    Exposure time: Lane1-4: 103 seconds; lane 5-6: 59 seconds

    All lanes: Western blot - Anti-TNFRSF10B antibody [RM2078] (ab323371) at 1/1000 dilution

    Lane 1: Parental HeLa (human cervical adenocarcinoma epithelial cell) whole lysate at 20 µg

    Lane 2: TNFRSF10B knockout HeLa whole lysate at 20 µg

    Lane 3: HT-1080 (human fibrosarcoma epithelial cell) whole lysate at 20 µg

    Lane 4: HepG2 (human hepatocelullar carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 5: Untreated HCT 116 (human colorectal carcinoma epithelial cell) whole lysate at 20 µg

    Lane 6: HCT 116 treated with 0.5uM doxorubicin for 24 hours whole 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.

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    FLRT3 is a glycoprotein of approximately 80-100 kDa and detected as a 75 kDa band after treated with Deglycosylation Peptide:N-glycosidase F (PNGase F).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-FLRT3 antibody [EPR29132-87] (ab323410) at 1/1000 dilution

    Lane 1: Untreated Huh7 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: Huh7 whole cell lysate treated with Protein Deglycosylation 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 1/20000 dilution

    Observed band size: 75-100 kDa, 36 kDa

    Exposure time: 15s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

    Negative control: Jurkat
    In Western blot, Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) ab244626 was shown to bind specifically to Clusterin. Target of interest was observed at 62 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in Clusterin knockout cell line (lane 2).
    The expression molecular weight observed is consistent with what has been described in the literature (PMID: 8373947).

    All lanes: Western blot - Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) (Anti-Clusterin antibody [EPR20729-240] - BSA and Azide free (Capture) ab244626) at 1/1000 dilution

    Lane 1: Wide-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: Clusterin knockout HeLa whole cell lysate at 20 µg

    Lane 3: T-47D (human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg

    Lane 4: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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: 62 kDa

    Exposure time: 52s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

    Exposure time:
    Lane 1-2: 180 seconds;
    Lane 3-4: 26 seconds

    All lanes: Western blot - Anti-PAI1 antibody [EPR21850-262] - BSA and Azide free (Detector) (Anti-PAI1 antibody [EPR21850-262] - BSA and Azide free (Detector) ab269409) at 1/1000 dilution

    Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: Human liver tissue lysate at 20 µg

    Lane 3: Mouse placenta tissue lysate at 20 µg

    Lane 4: Rat placenta 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: 45 kDa

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

    All lanes: Western blot - Anti-Caspase-3 antibody [EPR17664-28] - BSA and Azide free (Detector) (Anti-Caspase-3 antibody [EPR17664-28] - BSA and Azide free (Detector) ab281085) at 1/1000 dilution

    Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

    Lane 2: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

    Lane 3: 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

    Observed band size: 32 kDa

    Exposure time: 10s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    All lanes: Western blot - TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR19399] (TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR19399] ab197072) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

    Lane 2: 293T cells transfected with a human TrkA expression vector containing a His-tag, whole cell lysate at 20 µg

    Lane 3: 293T cells transfected with a human TrkB expression vector containing a His-tag, whole cell lysate at 20 µg

    Lane 4: 293T cells transfected with a human TrkC expression vector containing a His-tag, 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

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking buffer and dilution concentration: 5% NFDM/TBST.

    Lanes 1 - 4: Western blot - TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] (TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] ab229908) at 1/1000 dilution

    Lanes 1 - 4: Western blot - TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] - BSA and Azide free (TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR22298-67] - BSA and Azide free ab264070) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

    Lane 2: 293T cells transfected with a human TrkA expression vector containing a His-tag, whole cell lysate at 20 µg

    Lane 3: 293T cells transfected with a human TrkB expression vector containing a His-tag, whole cell lysate at 20 µg

    Lane 4: 293T cells transfected with a human TrkC expression vector containing a His-tag, 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 reducing conditions.

    Observed band size: 140 kDa

    Exposure time: 15s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Negative control: Daudi (PMID:32040549), KG-1a (PMID:32040549).

    The identity of the higher MW band at approximately 50 kDa is unknown.

    Samples are non-boiled as boiling may cause protein aggregation.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-GPCR GPRC5D antibody [EPR28376-41] (Anti-GPCR GPRC5D antibody [EPR28376-41] ab315808) at 1/1000 dilution

    Lane 1: RPMI-8226 (human plasmacytoma, myeloma B lymphocyte) whole cell lysate at 50 µg

    Lane 2: Daudi (human burkitt's lymphoma lymphoblast) whole cell lysate at 50 µg

    Lane 3: SK-MEL-2 (human skin malignant melanoma cell) whole cell lysate at 50 µg

    Lane 4: KG-1a (human bone marrow myeloblast) 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: 33 kDa, 36 kDa

    Exposure time: 180s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Negative control: human liver cancer.

    Samples are non-boiled as boiling may cause protein aggregation.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-GPCR GPRC5D antibody [EPR28376-41] (Anti-GPCR GPRC5D antibody [EPR28376-41] ab315808) at 1/1000 dilution

    Lane 1: SK-MEL-28 (human malignant melanoma cell) whole cell lysate at 60 µg

    Lane 2: Human liver cancer tissue lysate at 60 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Observed band size: 33 kDa, 36 kDa

    Exposure time: 180s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Low expression: skeletal muscle, colon, testis (PMID: 9490415).

    The identity of the bands higher 25 kDa are unknown.

    Samples are non-boiled as boiling may cause protein aggregation.

    The lanes 2-6 were developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-DAP12 antibody [EPR29151-41] (ab323318) at 1/1000 dilution

    Lane 1: Human lymph node tissue lysate at 20 µg

    Lane 2: Human skeletal muscle tissue lysate at 20 µg

    Lane 3: Human lung tissue at 20 µg

    Lane 4: Human placenta tissue lysate at 20 µg

    Lane 5: Human colon tissue lysate at 20 µg

    Lane 6: Human testis tissue lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG (HRP with minimal cross-reactivity with human IgG at 1/2000 dilution

    Developed using the ECL technique.

    Observed band size: 11 kDa, 12 kDa, 36 kDa

    Exposure time: 59s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression of Oncostatin-M is upregulated in response to LPS treatment (PMID: 12061841).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-Oncostatin M/OSM antibody [EPR26066-16] (ab323383) at 1/1000 dilution

    Lane 1: Untreated mouse lung tissue lysate at 50 µg

    Lane 2: Mouse lung treated with 1ug/ml LPS and 1ug/ml BFA for 16h 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: 28 kDa, 36 kDa

    Exposure time: 26s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Low expression: skeletal muscle.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-Oncostatin M/OSM antibody [EPR26066-16] (ab323383) at 1/1000 dilution

    Lane 1: Mouse spleen tissue lysate at 50 µg

    Lane 2: Mouse skeletal muscle 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: 28 kDa, 36 kDa

    Exposure time: 26s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 32641093).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-PITPnm 3 antibody [EPR28185-157] (ab323452) at 1/1000 dilution

    Lane 1: SK-OV-3 (human ovarian cancer epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

    Lane 2: SK-OV-3 transfected with siRNA specifically targeting PITPnm 3 whole cell 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: 106 kDa, 70 kDa, 36 kDa

    Exposure time: 10s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Negative control: Neuro-2a, 4T1, B16-F10.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 19415747). The band around 83 kDa is the full length and 69 kDa the alpha chain.

    The identity of the bands lower than 40 kDa are unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    Exposure time: Lanes 1-3: 180 seconds; Lanes 4-7: 37 seconds.

    All lanes: Western blot - Anti-HGF antibody [EPR27305-38] (ab323375) at 1/1000 dilution

    Lane 1: Mouse liver tissue lysate at 20 µg

    Lane 2: Mouse kidney tissue lysate at 20 µg

    Lane 3: Mouse spleen tissue lysate at 20 µg

    Lane 4: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

    Lane 5: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

    Lane 6: 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 7: B16-F10 (mouse skin melanoma 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: 83 kDa, 69 kDa, 36 kDa

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

    In Western blot, anti-WTAP antibody (Anti-WTAP antibody [EPR18744] ab195380) as pan control staining at 1/1000 dilution.

    In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

    All lanes: Western blot - Anti-WTAP (phospho S341) antibody [EPR27039-57] (Anti-WTAP (phospho S341) antibody [EPR27039-57] ab309350) at 1/1000 dilution

    Lane 1: Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate (untreated membrane) at 20 µg

    Lane 2: Wild-type HAP1 whole cell lysate (Alkaline phosphatase treated membrane) 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: 55 kDa

    Exposure time: 180s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The bands beneath the target band are likely to be degraded target fragments.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-TRF1 antibody [EPR28580-71] (ab323380) at 1/1000 dilution

    Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

    Lane 2: HeLa transfected with siRNA specifically targeting TRF1 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: 60 kDa, 36 kDa

    Exposure time: 48s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Low expression: THP-1, Huh7 (PMID: 26449829).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-PITPnm 3 antibody [EPR28185-157] (ab323452) at 1/1000 dilution

    Lane 1: HCC1937 (human mammary gland epithelial cell) whole cell lysate at 20 µg

    Lane 2: THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

    Lane 3: Huh7 (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 1/20000 dilution

    Observed band size: 106 kDa, 70 kDa, 36 kDa

    Exposure time: 37s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This antibody does not cross-react with mouse FLRT1 and FLRT2.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

    All lanes: Western blot - Anti-FLRT3 antibody [EPR29132-87] (ab323410) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

    Lane 2: 293T cells transfected with a mouse FLRT3 expression vector containing a GFP-tag, whole cell at 20 µg

    Lane 3: 293T cells transfected with a mouse FLRT1 expression vector containing a GFP-tag, whole cell lysate at 20 µg

    Lane 4: 293T cells transfected with a mouse FLRT2 expression vector containing a GFP-tag, 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: 75-100 kDa, 36 kDa

    Exposure time: 1s

  • Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602), expandable thumbnail

    Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID:7352261, PMID: 23027866 and PMID: 16020513).

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-Interferon beta antibody [RM1226] (Anti-Interferon beta antibody [RM1226] ab322112) at 1/1000 dilution

    Lane 1: A549 (human lung carcinoma epithelial cell) transfected with water instead of poly(I:C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, whole cell lysate at 20 µg

    Lane 2: A549 transfected with 400ng/ml poly(I:C) for 24 hours, 300ng/ml BFA was added to cells 4 hours after the start of the transfection, 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: 19 kDa, 22 kDa, 36 kDa

    Exposure time: 15s

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