A-20 Whole Cell Lysate

A-20 (B Lymphocyte; Reticulum Cell Sarcoma)

Tumor

RPMI 1640 Medium with 2 mM L-Glutamine & 10% FBS (Fetal Bovine Serum)

A-20 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 dodecyl sulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay.

The lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecyl sulfate, 0.01% bromophenol blue) containing 5% ß-mercaptoethanol.

One year from date of receipt

This lysate should be aliquoted in working amounts and may be stored for up to one month at -20°C <b>in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles.</b> For longer term storage, freeze working aliquots at -70°C.

This lysate is for use in Western blotting, 10 µg to 20 µg per lane is recommended for mini gel.