3T3 (Balb) 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 3T3 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% β-mercaptoethanol.
|Source||BALB/3T3 clone A31 (Mouse embryo)|
|Concentration||Product is supplied at a concentration of 2 mg/ml.|
|Reconstitution||During shipment, small volumes will occasionally become entrapped in the seal of the product vial. For products with volumes of 200 μl 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.|
|Stability & Storage||Store at 4°C for continuous use. Store at -20°C for three months or at -70°C for up to one year. Avoid repeated freezing and thawing (store working aliquots at -70°C). Under proper storage conditions the shelf life is 6 months from the date of receipt.|