GAPDH mouse mAb #50202

GAPDH Antibody

Background: Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. GAPDH is constitutively expressed in almost all tissues at high levels, therefore antibodies against GAPDH are useful as loading controls for Western Blotting. Some physiological factors, such as hypoxia and diabetes, increase GAPDH expression in certain cell types.

Formulation: 100uL of mouse monoclonal IgG in 1mg/mL of 10mM Tris-HCl, pH 8.0, 150mM NaCl, 25% glycerol, 0.02% sodium azide.

Storage: Store the product at -20°C. Product is stable for about 6 weeks at 2-8°C as an undiluted liquid. Avoid repeated freezing and thawing.

Note: For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Immunoblot Protocol

Perform SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on samples and transfer the proteins to a PVDF membrane. Block the blotted PVDF membrane in freshly prepared PBS containing 5% milk with for 1 hour at room temperature with constant agitation. Incubate the PVDF with the recommended dilution of GAPDH antibody(#50202) diluted in freshly prepared 5% milk for 1 hour at room temperature or overnight with agitation at 2-8°C. Wash the PVDF 3 times with PBS. Incubate the PVDF in the secondary reagent of choice (a goat anti-mouse secondary antibody conjugated to HRP, Catalog # K009, 1:3000 dilution was used) in 5% milk for 1 hour with agitation at room temperature. Wash the PVDF 3-5 times with PBS. Incubate the PVDF in ECL substrate for 1 min and cover the membrane by PE film. In the darkroom, expose the PVDF to Autoradiography film for 5s~ 5 min to visualize results.

本产品仅供研究使用/For Research Use Only.