Description |
Lung tissue 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 ug/ml of aprotinin, 5 ug/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product 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% beta-mercaptoethanol. |
Reconstitution |
During shipment, small volumes of product will occasionally become entrapped in the seal of the product vial. For lysates with volumes of 200 uL or less, we recommend gently tapping the vial on a hard surface or briefly centrifuging the vial in a tabletop centrifuge to dislodge any liquid in the container's cap. |
Storage and Stability |
It is supplied as whole cell lysate, 100 ug per vial at 2.0 mg/ml, in 1 x SDS sample buffer containing 5% beta-mercaptoethanol. Storage at -20C is stable for three months, -80C for one year. |