Overview
- Product nameAnti-GAPDH antibodySee all GAPDH primary antibodies ...
- DescriptionRabbit polyclonal to GAPDH
- Tested applicationsIP, ELISA, WB, IHC-Fr, ICC/IF, Flow Cyt more details
- Species reactivityReacts with: Mouse, Rat, Chicken, Human, Saccharomyces cerevisiae, Xenopus laevis, Schizosaccharomyces pombe, African Green Monkey
- Immunogen
Full length native protein from human erythrocytes.
- Positive controlWB: CHO and HEK 293 cell lysate ICC: U2OS cells
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: PBS, pH 7.4 -
Concentration information loading... - PurityProtein A purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab9485 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| IP | IP: 1/250. |
| ELISA | ELISA: 1/2500 - 1/5000. |
| WB | WB: 1/2500. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa). Some customers have experienced that milk significantly decreases the signal in Wb compared to BSA. In-house we use BSA. |
| IHC-Fr | IHC-Fr: 1/250. |
| ICC/IF | ICC/IF: Use a concentration of 5 µg/ml. |
| Flow Cyt | Flow Cyt: Use at an assay dependent dilution. |
Target
- FunctionHas 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.
- PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
- Sequence similaritiesBelongs 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 localizationCytoplasm > 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.
-
Database links
- Entrez Gene: 374193 Chicken
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 24383 Rat
- Entrez Gene: 685186 Rat
- Entrez Gene: 380259 Xenopus laevis
- Omim: 138400 Human
- SwissProt: P00356 Chicken
- SwissProt: P04406 Human
- SwissProt: P16858 Mouse
- SwissProt: P04797 Rat
- SwissProt: P51469 Xenopus laevis
- Unigene: 544577 Human
- Unigene: 592355 Human
- Unigene: 598320 Human
- Unigene: 304088 Mouse
- Unigene: 309092 Mouse
- Unigene: 317779 Mouse
- Unigene: 343110 Mouse
- Unigene: 392463 Mouse
- Unigene: 392480 Mouse
- Unigene: 414470 Mouse
- Unigene: 458138 Mouse
- Unigene: 458416 Mouse
- Unigene: 475698 Mouse
- Unigene: 129558 Rat
- Unigene: 91450 Rat
- Unigene: 995 Xenopus laevis
see all
Target information above from: UniProt accession
P04406
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
.
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Alternative names
- Aging associated gene 9 protein antibodyG3P_HUMAN antibodyG3PD antibody
- G3PD antibodyG3PDH antibodyGAPD antibodyGAPDH antibodyGAPDH antibodyGlyceraldehyde 3 phosphate dehydrogenase antibodyGlyceraldehyde 3 phosphate dehydrogenase liver antibodyGlyceraldehyde 3 phosphate dehydrogenase muscle antibodyGlyceraldehyde-3-phosphate dehydrogenase antibodyMGC88685 antibodyOCAS p38 component antibodyOCT1 coactivator in S phase 38 KD Component antibodyPeptidyl-cysteine S-nitrosylase GAPDH antibody
see all
Anti-GAPDH antibody images
-
Western blot - GAPDH antibody - Loading Control (ab9485)All lanes : Anti-GAPDH antibody (ab9485) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : A431 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : 293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 37 kDa
Observed band size : 40 kDa (why is the actual band size different from the predicted?)
Exposure time : 1 second -
Immunocytochemistry/ Immunofluorescence - GAPDH antibody (ab9485)This image is courtesy of an anonymous Abreviewab9485 staining GAPDH (green) in Rat bone marrow cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X 100 and blocked with 5% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/250 in PBS) for 12 hours at 4°C. An Alexa Fluor®488-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody. Nuclei were stained with DAPI. -
Western blot - Anti-GAPDH antibody (ab9485)
Predicted band size : 37 kDa -
Immunocytochemistry/ Immunofluorescence - GAPDH antibody (ab9485)ICC/IF image of ab9485 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab9485, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). -
Western blot - GAPDH antibody (ab9485)All lanes : Anti-GAPDH antibody (ab9485) at 1/2500 dilution
Lane 1 : Lysate prepared from human Huh-7 cells at 2 µg
Lane 2 : Lysate prepared from human Huh-7 cells at 20 µg
Secondary
HRP-conjugated sheep polyclonal to rabbit IgG at 1/20000 dilution
Performed under reducing conditions.
Predicted band size : 37 kDa
Observed band size : 40 kDa (why is the actual band size different from the predicted?)
Exposure time : 5 minutesThis image is a courtesy of Anonymous Abreview
-
Western blot - GAPDH antibody (ab9485)
Predicted band size : 37 kDa
This GAPDH antibody works well with an S. cerevisiae lysate in Western blotting.A band can be seen down to a dilution of 1/5000.
A: 1:1000, 25 ug lysate
B: 1:1000, 50 ug lysate
C: 1:2000, 25 ug lysate
D: 1:2000, 50 ug lysate
E: 1:5000, 25 ug lysate
F: 1:5000, 50 ug lysate
This picture was generously supplied by Dr Allison Robb of the MRC-LMB, Cambridge, UK.
References for Anti-GAPDH antibody (ab9485)
This product has been referenced in:
- Teh MT et al. FOXM1 Induces a Global Methylation Signature That Mimics the Cancer Epigenome in Head and Neck Squamous Cell Carcinoma. PLoS One 7:e34329 (2012). WB ; Human . Read more (PubMed: 22461910) »
- Castella M et al. Origin, functional role, and clinical impact of Fanconi anemia FANCA mutations. Blood 117:3759-69 (2011). WB ; Human . Read more (PubMed: 21273304) »
