Recombinant Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16891] to GAPDH - Loading Control
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Chicken, Human, Zebrafish, African green monkey, Xenopus tropicalis
Related conjugates and formulations
Overview
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Product name
Anti-GAPDH antibody [EPR16891] - Loading Control
See all GAPDH primary antibodies -
Description
Rabbit monoclonal [EPR16891] to GAPDH - Loading Control -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Chicken, Human, Zebrafish, African green monkey, Xenopus tropicalis
Predicted to work with: Rabbit, Fish -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, UMNSAH/DF-1, Jurkat, COS-1, RAW 264.7 and PC-12 whole cell lysates; Human fetal brain and heart lysates; Xenopus(X. tropicalis) muscle lysate; Zebrafish lysate; Mouse kidney and spleen lysates; Rat brain lysate. IHC-P: Human transitional cell carcinoma of bladder, Mouse spleen and Rat spleen tissues. ICC/IF: HeLa cells. Flow: Jurkat cells. IP: HeLa whole cell extract
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16891 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-GAPDH antibody [EPR16891] - BSA and Azide free (ab199553)
- 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)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
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- Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
- Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
- Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
- Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab181602 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/180.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (19) |
1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (3) |
1/500.
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IP |
1/60.
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Notes |
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Flow Cyt (Intra)
1/180. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/500. |
IP
1/60. |
Target
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Function
Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. 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). 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. -
Pathway
Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5. -
Sequence similarities
Belongs to the glyceraldehyde-3-phosphate dehydrogenase family. -
Post-translational
modificationsS-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
ISGylated. -
Cellular localization
Cytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions. - Information by UniProt
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Database links
- Entrez Gene: 374193 Chicken
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 100009074 Rabbit
- Entrez Gene: 24383 Rat
- Entrez Gene: 685186 Rat
- Entrez Gene: 317743 Zebrafish
see all -
Alternative names
- 38 kDa BFA-dependent ADP-ribosylation substrate antibody
- aging associated gene 9 protein antibody
- Aging-associated gene 9 protein antibody
see all
Images
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All lanes : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution
Lane 1 : Mouse kidney lysates
Lane 2 : Mouse spleen lysates
Lane 3 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 5 : Rat brain lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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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 (ab150077) at a dilution of 1/400. Nucleus was counter-stained with DAPI (blue). ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with 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 (ab150120; 1/500) was used. For negative control 2 primary antibody (ab7291; 1/500) and secondary antibody (ab150077; 1/400) was used.
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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.
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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.
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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.
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All lanes : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/50000 dilution
Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : Xenopus tropicalis muscle lysates
Lane 3 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates
Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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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.
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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.
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All lanes : 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
Lane 2 : Zebrafish lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution
Lane 1 : Human fetal brain lysates
Lane 2 : Human fetal heart lysates
Lysates/proteins at 10 µg per lane.
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 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (1571)
ab181602 has been referenced in 1571 publications.
- Yan Y et al. The circular RNA hsa_circ_0001394 promotes hepatocellular carcinoma progression by targeting the miR-527/UBE2A axis. Cell Death Discov 8:81 (2022). WB ; Human . PubMed: 35210429
- Liu X et al. m6A methylated EphA2 and VEGFA through IGF2BP2/3 regulation promotes vasculogenic mimicry in colorectal cancer via PI3K/AKT and ERK1/2 signaling. Cell Death Dis 13:483 (2022). WB ; Human . PubMed: 35595748
- Liu F et al. LncRNA LIFR-AS1 overexpression suppressed the progression of serous ovarian carcinoma. J Clin Lab Anal N/A:e25470 (2022). WB ; Human . PubMed: 35778954
- Guo S et al. Ligustilide alleviates the insulin resistance, lipid accumulation, and pathological injury with elevated phosphorylated AMPK level in rats with diabetes mellitus. J Recept Signal Transduct Res 41:85-92 (2021). PubMed: 32643505
- Lu W et al. Reprogramming immunosuppressive myeloid cells facilitates immunotherapy for colorectal cancer. EMBO Mol Med 13:e12798 (2021). PubMed: 33283987