Description:
A freshly-harvested Sprague Dawley rat heart is flash-frozen in liquid nitrogen within 30
seconds of removal and stored frozen until lysis. The tissue is ground and thoroughly
homogenized at 4°C with a freshly prepared modified Radio Immuno Precipitation Assay
(RIPA) buffer containing strong protease and phosphatase inhibitor cocktails. The final
homogenate is incubated with DNase then clarified by centrifugation at 10,000 g at 4°C
and stored at -80°C. Each lot is quality controlled by determining the protein
concentration using a BCA assay and analyzed with SDS-PAGE using Coomassie blue
for visualization. Sharp, clear protein bands indicate a quality protein lysate.
Formulation:
Liquid in 0.1M Tris-HCl, 0.15M NaCl, 0.1% SDS, 0.5% Na Deoxycholate, 1% Triton X -100, pH 7.4
Storage/Stability:
Store at -80°C for up to 1 year. Upon initial thaw, prepare working aliquots and store at -80°C. Avoid repeated freeze-thaw cycles.
