Rat brain tissue lysate (denatured)
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Specification
Product Description
Whole tissue lysate (denatured).
Tissue
Brain
Host
Rat
Preparation Method
Tissue 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 adjusted to 3.75 mg/ml, and then mixed with 5X Sample Buffer to become final 3 mg/ml in 1X Sample 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.
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.
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Applications
Western Blot
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