Western blot analysis of GAPDH was performed by loading 50ug of various cell lysates per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a biotinylated GAPDH monoclonal antibody (Product # MA5-15738-BTIN) at a dilution of 1:1000 for 1 hour at room temperature, followed by Streptavidin-HRP (Product # 21126) at a dilution of 1:20,000 for 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).
|Tested species reactivity||Bacteria, Chicken, Hamster, Human, Insect, Mouse, Rabbit, Rat, Yeast|
|Host / Isotype||Mouse / IgG1|
|Storage buffer||PBS with proprietary stabilizer|
|Contains||0.02% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||2 ug/test|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:500|
|Western Blot (WB)||1:500 - 1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA5-15738-BTIN has successfully been used for Western blot, FACS, ELISA, IHC (P) and ICC/IF applications.
Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is an enzyme involved in glycolysis. In addition to this role, it is involved in other cellular processes ranging from membrane fusion, neuronal apoptosis to cancer. GAPDH is expressed at high levels in most tissues and is therefore useful as protein loading control in western blot analysis. Additionally, the antigen is conserved in most eukaryotes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.