Anti-GAPDH antibody (ab22555)
- Product nameAnti-GAPDH antibodySee all GAPDH primary antibodies ...
- DescriptionRabbit polyclonal to GAPDH
- Tested applicationsICC/IF, WB more details
- Species reactivityReacts with: Mouse, Rat, Chicken, Human, Baboon
Predicted to work with: Sheep, Rabbit, Horse, Guinea pig, Cow, Cat, Dog, Turkey, Pig, Duck
Synthetic peptide: DAPMFVMGVNHFKYA , corresponding to amino acids 126-140 of Human GAPDH.
- Positive control
- Rat heart protein extract. This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: MCF7
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPreservative: 0.05% Sodium Azide
Constituents: PBS, 1mg/ml BSA
- 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 ab22555 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.|
|WB||WB: Use a concentration of 0.1 µg/ml. Detects a band of approximately 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: 493876 Cat
- Entrez Gene: 374193 Chicken
- Entrez Gene: 281181 Cow
- Entrez Gene: 403755 Dog
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 396823 Pig
- Entrez Gene: 100009074 Rabbit
- Entrez Gene: 24383 Rat
- Entrez Gene: 685186 Rat
- Entrez Gene: 443005 Sheep
- Omim: 138400 Human
- SwissProt: Cat
- SwissProt: P00356 Chicken
- SwissProt: P10096 Cow
- SwissProt: Q28259 Dog
- SwissProt: P70685 Guinea pig
- SwissProt: P04406 Human
- SwissProt: P16858 Mouse
- SwissProt: P00355 Pig
- SwissProt: P46406 Rabbit
- SwissProt: P04797 Rat
- SwissProt: Q28554 Sheep
- 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
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Anti-GAPDH antibody images
Anti-GAPDH antibody (ab22555) at 0.1 µg/ml + Rat heart protein extract
Performed under reducing conditions.
Observed band size : 36 kDa (why is the actual band size different from the predicted?)
All lanes : Anti-GAPDH antibody (ab22555) at 1/5000 dilution
Lane 1 : Mouse Tissue Lysate- Whole brain
Lane 2 : Mouse Tissue Lysate- Whole brain
Lane 3 : Mouse Tissue Lysate- Whole brain
Lane 4 : Mouse Tissue Lysate- Whole brain
Lane 5 : Mouse Tissue Lysate- Whole brain
Lane 6 : Mouse Tissue Lysate- Whole brain
Lane 7 : Mouse Tissue Lysate- Whole brain
Lane 8 : Mouse Tissue Lysate- Whole brain
Lysates/proteins at 50 µg per lane.
This image is courtesy of an anonymous Abreview
All Lanes: ab22555 (1/15000) staining GAPDH in Mouse brain whole tissue lysate (50 µg).
Blocking Step: 5% Milk for 1 hour at 20°C.
Secondary Antibody: HRP-conjugated Goat anti-rabbit IgG polyclonal (1/30000).
ab22555 stained MCF-7 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 ab22555 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.
References for Anti-GAPDH antibody (ab22555)
This product has been referenced in:
- van Blitterswijk M et al. Characterization of FUS mutations in amyotrophic lateral sclerosis using RNA-Seq. PLoS One 8:e60788 (2013). WB ; Human . Read more (PubMed: 23577159) »
- Finkensieper A et al. The 5-lipoxygenase pathway regulates vasculogenesis in differentiating mouse embryonic stem cells. Cardiovasc Res 86:37-44 (2010). Read more (PubMed: 19965960) »