Description:
Rat spleen tissue lysate was prepared by homogenization in modified RIPA buffer (150
mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1
mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1%
sodium dodecylsulfate, 5 µg/ml of aprotinin, 5 µg/ml of leupeptin). Tissue and cell debris
was removed by centrifugation. Protein concentration was determined with Bio-Rad
protein assay. The tissue lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM
Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue)
containing 50 mM DTT.
Application notes:
Centrifuge vial before use. Rat spleen tissue lysate is ready to load on SDS-PAGE for Western blotting, 10-20 µg per lane is recommended for mini gel. The optimal concentration should be determined for each specific application.