HT-29 (Human Colorectal Adenocarcinoma)
ATCC Medium (McCoy’s 5a Medium with 1.5 mM L-Glutamine, and 10% FBS)
HT-29 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. 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 dodecylsulfate, 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 in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. For longer term storage, freeze working aliquots at -70°C.
Applications and Suggested Working Dilutions
This lysate is for use in Western blotting, 10 µg to 20 µg per lane is recommended for mini gel.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.