This lysate is prepared as a positive control for separation by SDS-PAGE and subsequent Western blot analysis. Lysates are prepared in denaturing buffer WITHOUT reducing agents (i.e. 2-mercaptoethanol (BME) or dithiothreitol (DTT)). If reducing conditions are desired, add a reducing agent prior to heating. Heat lysate to 95 degrees C for 5 minutes and rapidly cool. The recommended loading amount is 0.01-0.03 mg per lane.
Packaging, Storage & Formulations
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Standard RIPA buffer including protease inhibitors and 0.1% SDS diluted in 1 X lithium dodecyl sulfate (LDS) sample buffer with bromophenol blue and 40-70% glycerol, pH 8.4. Denaturing agents have not been added.
Lysate Details for HeLa
HeLa whole cell lysate was lysed in modified RIPA buffer (150 mM sodium chloride, 25 mM Tris-HCl, pH 7.6, 1% Triton X-100, 1% sodium deoxycholic acid, 1% sodium dodecyl sulfate, 1 mM phenyl methyl sulfonyl fluoride, 1 ug/ml of aprotinin, 1 ug/ml of leupeptin, 1ug/ml of pepstatin). Protein concentration was determined with Bio-Rad protein assay.
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed
for 6 months from date of receipt.
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