Mouse spleen tissue lysate (non-denatured)
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Specification
Product Description
Whole tissue lysate (non-denatured).
Tissue
Spleen
Host
Mouse
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 5 mg/ml.
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
Use it directly for immuno-precipitation, or heat lysate with SDS gel loading buffer to 95°C for 5 minutes followed by rapid cooling for western blot application. If dissociating conditions are required, add reducing agent prior to heating.
Storage Buffer
In modified RIPA Lysis Buffer.
Storage Instruction
Store at -80°C. Aliquot to avoid repeated freezing and thawing.
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Applications
Western Blot
Immunoprecipitation
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