Description:
Human HT-29 Cell Line 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 flouride + 5 µg/ml aprotinin + 5 µg/ml leupeptin).
Cell debris was removed by centrifugation. Protein concentration was determined with
protein assay. Lysate was boiled for 5 minutes in 1x SDS sample buffer (50 mM Tris-HCl
pH 6.8 + 12.5% Glycerol + 1% SDS + 0.01% bromophenol blue) containing 5% betamercaptoethanol.
Formulation:
Liquid in 1x SDS sample buffer containing 5% beta-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. HT-29 Cell Lysate is ready to load on SDS-PAGE for Western Blotting. It is recommended to load 10 - 20 µg per lane for a mini gel. The optimal concentration should be determined by the user for each specific application.
