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Western blot - GAPDH antibody - Loading Control (ab9485)
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Western blot - GAPDH antibody (ab9485)
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Immunocytochemistry/ Immunofluorescence - GAPDH antibody (ab9485)
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Product Name
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GAPDH antibody
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See all GAPDH - Loading Control antibodies (10)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to GAPDH
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Immunogen
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Full length native protein from human erythrocytes.
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Reacts with
(species key)
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Hu, Ms, Rat, Chk, Xl
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Tested applications
(see key)
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ELISA, Flow Cyt, ICC/IF, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ELISA: 1/2500 - 1/5000.
Flow Cyt: Use at an assay dependent dilution.
ICC/IF: Use at a concentration of 5 µg/ml.
IP: 1/250.
WB: 1/2500. Detects a band of approximately 40 kDa (predicted molecular weight: 37 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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WB: CHO and HEK 293 cell lysate
ICC: U2OS cells
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Cellular localization
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Cytoplasmic
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Research areas
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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
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Relevance
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Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. Besides its functioning as a glycolytic enzyme in cytoplasm, recent evidence suggest that mammalian GAPDH is also involved in a great number of intracellular proceses such as membrane fusion, microtubule bundling, phosphotransferase activity, nuclear RNA export, DNA replication, and DNA repair. During the last decade a lot of findings appeared concerning the role of GAPDH in different pathologies including prostate cancer progression, programmed neuronal cell death, age-related neuronal diseases, such as Alzheimer's and Huntington's disease. GAPDH is constitutively expressed in almost all tissues at high levels, therefore becoming the marker of choice when a loading control in Western blotting is required. Some physiological factors, such as hypoxia and diabetes, increase GAPDH expression in certain cell types.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Protein A purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.90 - 1.00 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this GAPDH antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for GAPDH antibody images, references, products related to ab9485 and other tools.
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GAPDH antibody images:
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 Western blot - GAPDH antibody - Loading Control (ab9485)
All lanes : 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 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
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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.
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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).
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References for GAPDH antibody (ab9485)
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This product has been used in: (two most recent references)
Andrieu C et al. Heat shock protein 27 confers resistance to androgen ablation and chemotherapy in prostate cancer cells through eIF4E. Oncogene : (2010). WB; Human. PubMed: 20101233
Ramseger R et al. Transmembrane form agrin-induced process formation requires lipid rafts and the activation of Fyn and MAPK. J Biol Chem 284:7697-705 (2009). WB; Chicken. PubMed: 19139104
See all 87 publication references for this product.
If you publish research using ab9485 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to GAPDH
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GAPDH antibody - more information
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Customer reviews (feedback) regarding GAPDH antibody |
Customer FAQs regarding GAPDH antibody |
Protocols for GAPDH antibody |
Price and availability of products related to GAPDH antibody |
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GAPDH antibody - related products:
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GAPDH antibody - other tools:
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Contact Abcam with a Technical Enquiry about GAPDH antibody |
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All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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