3T3 NIH Whole Cell Lysate

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.