Anti-GAPDH antibody (ab70699)
- Product nameAnti-GAPDH antibodySee all GAPDH primary antibodies ...
- DescriptionRabbit polyclonal to GAPDH
- Tested applicationsICC/IF, IHC-P, WB more details
- Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Chicken, Guinea pig, Turkey, Pig, Xenopus laevis, Chimpanzee, Reptiles, Rhesus monkey, Gorilla
Synthetic peptide, corresponding to a region between residues 250-300 of Human GAPDH. (NP_002037.2).
- Positive control
- Whole cell lysate from HeLa and mouse NIH3T3 cells. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal skeletal muscle. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPreservative: 0.09% Sodium Azide
Constituents: 0.1% BSA, Tris buffered saline
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
Our Abpromise guarantee covers the use of ab70699 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||ICC/IF: Use a concentration of 5 µg/ml.|
|IHC-P||IHC-P: Use a concentration of 5 µg/ml.|
|WB||WB: 1/2000 - 1/10000. Detects a band of approximately 38 kDa (predicted molecular weight: 36 kDa).|
- 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.
modificationsS-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
- 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.
- Entrez Gene: 374193 Chicken
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 396823 Pig
- Entrez Gene: 24383 Rat
- Entrez Gene: 685186 Rat
- Entrez Gene: 380259 Xenopus laevis
- Omim: 138400 Human
- SwissProt: P00356 Chicken
- SwissProt: P70685 Guinea pig
- SwissProt: P04406 Human
- SwissProt: P16858 Mouse
- SwissProt: P00355 Pig
- 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
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Anti-GAPDH antibody images
ab70699 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab70699 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
All lanes : Anti-GAPDH antibody (ab70699) at 0.04 µg/ml
Lane 1 : Whole cell lysate from HeLa cells at 50 µg
Lane 2 : Whole cell lysate from HeLa cells at 15 µg
Lane 3 : Whole cell lysate from HeLa cells at 5 µg
Lane 4 : Whole cell lysate from NIH 3T3 cells at 50 µg
Predicted band size : 36 kDa
Observed band size : 38 kDa (why is the actual band size different from the predicted?)
Exposure time : 10 seconds
IHC image of GAPDH staining in Human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab70699, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
References for Anti-GAPDH antibody (ab70699)
This product has been referenced in:
- Charron G et al. Prenylome profiling reveals S-farnesylation is crucial for membrane targeting and antiviral activity of ZAP long-isoform. Proc Natl Acad Sci U S A 110:11085-90 (2013). Read more (PubMed: 23776219) »
- Yount JS et al. S-Palmitoylation and Ubiquitination Differentially Regulate Interferon-induced Transmembrane Protein 3 (IFITM3)-mediated Resistance to Influenza Virus. J Biol Chem 287:19631-41 (2012). WB ; Human . Read more (PubMed: 22511783) »