Details
Cell Line
NIH/3T3 (embryo)
Tissue Type
Fibroblast from NIH Swiss mouse embryos
Growth Media
DMEM & 10% fetal bovine serum
Preparation
Mouse NIH/3T3 cell 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 dodecyl sulfate (SDS), 1 mM sodium ethylenediaminetetraacetate, 1 mM phenylmethyl-sulfonyl fluoride, 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% SDS, 0.01% bromophenol blue) containing 5% β-mercaptoethanol.
Host
Mouse
Concentration
2.0 mg/ml
Formulation
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 dodecyl sulfate, 0.01% bromophenol blue) containing 5% ß-mercaptoethanol.
Expiration Date
One year from date of receipt
Storage
This whole cell lysate is stable for three months at -20°C or at –70°C for up to one year.
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.