HT29

HT29 cells that produce carcinoembryonic antigen (CEA) and are human intestinal epithelial cells which produce the secretory component of immunoglobulin A (IgA). These cells are used for tumourigenicity studies. The lysate is a good positive control for keratin 18, keratin 20 (several antibodies in our catalog), the protooncogene cyclin D1 (several antibodies in our catalog). HT 29 cell lysate is prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylene diamine tetraacetic acid, 1 mM phenylmethylsulfonyl flouride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecyl sulfate, 5 µg/ml of aprotinin, and 5 µg/ml of leupeptin). Cell debris is removed by centrifugation. Protein concentration is determined via Bio-Rad protein assay. The cell lysate is boiled for five minutes in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1%sodium dodecylsulfate, and 0.01% bromophenol blue) containing 5% b-mercaptoethanol.