Description:
Rat colon tissue lysate was prepared by homogenization in modified RIPA buffer (50 mM
Tris-HCl, pH 7.4 + 1% Triton X-100 + 0.2% sodium deoxycholate + 0.2% sodium
dodecylsulfate (SDS) + 1 mM sodium ethylenediaminetetraacetate + 1 mM
phenylmethylsulfonyl fluoride + 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 lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM
Tris -HCl, pH 6.8 + 12.5% glycerol + 1% SDS + 0.01% bromophenol blue) containing 5%
β-mercaptoethanol.
Formulation:
Colon tissue lysate, 100 µg per vial, at 2.0 mg/ml, in 1 x SDS sample buffer containing 5% β-mercaptoethanol.
Storage/Stability:
Store at 2-8°C for continuous use. Store in working aliquots at -80°C for up to 1 year. Avoid repeated freeze-thaw cycles.
Application notes:
Centrifuge vial before use. Rat colon tissue lysate is ready to load on SDS-PAGE for Western blotting. It is recommended to load 10 µg to 20 µg per lane for mini gel.
