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Products:Neuroscience >> Neurology process >> Neurodegenerative disease >> Huntington's disease
Overview
Product name
Anti-GAPDH antibody
See all GAPDH products (9) ...
Description
Rabbit polyclonal to GAPDH
Tested applications
IP, ELISA, WB, IHC-Fr, ICC/IF, Flow Cytmore details
Cross reactivity
Reacts with
Mouse, Rat, Chicken, Human, Saccharomyces cerevisiae, Xenopus laevis
Immunogen
Full length native protein from human erythrocytes.
Positive control
WB: CHO and HEK 293 cell lysate ICC: U2OS cells
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Protein A purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Cancer >> Cancer Metabolism >> Metabolic signaling pathway >> Metabolism of carbohydrates
Signal Transduction >> Metabolism >> Energy Metabolism
Isotype/Loading Controls >> Loading Controls >> GAPDH
Neuroscience >> Neurology process >> Neurodegenerative disease >> Alzheimer's disease >> Other
Neuroscience >> Neurology process >> Neurodegenerative disease >> Huntington's disease
Applications
Show applications key
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.
IP: 1/250
ELISA: 1/2500 - 1/5000.
WB: 1/2500Detects 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: 1/250
ICC/IF: Use a concentration of 5 µg/ml
Flow Cyt: Use at an assay dependent dilution.
Target
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
modifications
S-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.
Target information above from: UniProt accessionP04406
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Aging associated gene 9 protein antibody
- G3P_HUMAN antibody
- G3PD antibody
see all
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
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)
ab9485 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.
This image is courtesy of an anonymous Abreview
Western blot - Anti-GAPDH antibody (ab9485)
Predicted band size : 37 kDa
Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) (ab119220) with the
Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623) .
20ug of Lysate per lane and detection using ab9485 diluted to 1ug/ml.
Lane 1: Hela cell lysate
Lane 2: Jurkat cell lysate
Lane 3: A431 cell lysate
Lane 4: HEK293 cell lysate
Lane 5: HepG2 cell lysate
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 minutes
This 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:
- Wood MAet al. Using dihydropyridine-release strategies to enhance load effects in engineered human bone constructs. Tissue Eng 12:2489-97 (2006).Read more (PubMed: 16995782) »
- Castella Met al. Origin, functional role, and clinical impact of Fanconi anemia FANCA mutations. Blood 117:3759-69 (2011). WB; Human.Read more (PubMed: 21273304) »
See all 92 publications for this product
Publishing research using ab9485? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"