NIH-3T3 embryonic cells grown to 85% confluency were harvested, homogenized in modified RIPA buffer (150 mM NaCl, 50 mM Tris-Cl, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, 1 mM EDTA, 1 mM PMSF, 5 μg/ml Pepstatin A, 2 μg/ml Leupeptin, 1 mM sodium orthovanadate, 1 mM sodium fluoride.), and cleared by centrifugation. Total protein concentration of the lysate was determined using BCA protein assay (BSA was used as a standard). Lysate was mixed with Laemmli Sample Buffer containing reducing reagent and was boiled for 5 minutes. Lysate is supplied as 200 μg in 100 μl per vial (2.0 mg/ml total protein concentration) in 1X Laemmli sample buffer.
1x SDS sample buffer, 5% β-mercaptoethanol
Store as concentrated solution. Centrifuge briefly prior to opening vial. Aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.
1 mg/ml (Please refer to the vial label for the specific concentration.)
Whole Cell Lysate
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.