Caco-2 Whole Cell Lysate

Caco-2 (Human Colorectal Adenocarcinoma)

ATCC Medium (Minimum Essential Medium (Eagle) with 2mM L-Glutamine, 1.5 g/L Sodium Bicarbonate, 0.1 mM non-Essential Amino Acids, 1.0 mM Sodium Pyruvate, and 20% FBS.)

Caco-2 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 2-8°C. Avoid Repeated Freeze Thaw Cycles.</b> For longer term storage, freeze working aliquots at -70°C <b> in a manual defrost freezer.

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