Western blot analysis of Mouse Heart Tissue Control Extracts using Phospho-Troponin I (Cardiac) (Ser23/24) Antibody #4004.
Western blot analysis of Mouse Heart Tissue Control Extracts using Troponin I Antibody #4002.
Western blot analysis of Mouse Heart Tissue Control Extracts using Myosin Light Chain 2v (D5I1C) (Cardiac Isoform) Rabbit mAb (Rodent Specific) #12975.
CST recommends loading 10-20 μl extract per mini-gel lane. Boil for 2 minutes prior to use.
Supplied in SDS Sample Buffer. Store at –20°C, or at –80°C for long-term storage.
The extract is prepared from whole heart tissue of adult mice, and is intended for use as a positive control in western blotting applications. The protein concentration is 2 mg/ml.
The extract was prepared from whole tissue by homogenization in 1X RIPA buffer (#9806, 10X) (20 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 1 mM sodium EDTA, 1 mM EGTA, 1 μg/ml leupeptin, 1 mM β-glycerophosphate, 1 mM sodium orthovanadate, 2.5 mM sodium pyrophosphate), 1 mM PMSF (#8553, 34.84 mg), 1X Protease/Phosphatase Inhibitor Cocktail (#5872, 100X).
The extract was subsequently sonicated and insoluble cell debris removed by centrifugation. The extract was then boiled for 5 min in 1X SDS sample buffer + DTT (#7722, Blue Loading Buffer Pack) (62.5 mM Tris-HCl pH 6.8, 10% glycerol, 2% SDS, 0.01% bromophenol blue, 42 mM DTT) to denature the proteins.
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