Description
Cell or tissue lysates are a useful control for the western blotting application. A source that can be rich in endogenously expressed proteins, this lysate is convenient to use and provided in a denatured format allowing for immediate loading onto an SDS-PAGE gel.
This lysate was routinely tested by western blot analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
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Preparation and Storage
Store undiluted at -20°C.Do not dilute or boil.NIH 3T3 cells were serum starved overnight and then incubated at room temperature for 10 minutes in a final concentration of 1 mM sodium vanadate. PDGF was then added at a final concentration of 5 ng/ml and cells were incubated at 37°C for 30 minutes.Western blots of NIH 3T3 cell lysates treated with vanadate and PDGF have been reported to show a shift in SHC mobility in addition to phosphorylation of PKBα/Akt and Stat3 as compared to untreated control cells.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
