Products

Product Description

Whole cell lysate (denatured).

Tissue

Macrophage

Host

Mouse

Preparation Method

Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined with Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The lysate was mixed in 5X Sample Buffer to become final 1X Sample Buffer as Storage Buffer. The lysate was heated at 95°C for 5 min, and cooled rapidly.

Lysis Buffer

Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.

Quality Control Testing

12.5% SDS-PAGE Stained with Coomassie Blue.

SDS-PAGE Gel

Recommend Usage

The lysate is ready to load on SDS-PAGE for Western blotting.

Storage Buffer

In ready-to-use 1X Sample Buffer (50 mM Tris-HCl, 2% SDS, 10% glycerol, 300 mM 2-mercaptoethanol, 0.01% Bromophenol blue).

Storage Instruction

Store at -80°C. Aliquot to avoid repeated freezing and thawing.