|Tel :||01904 431402||Bioquote Limited|
The Raylor Centre
York, YO10 3DW
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|Int Tel :||00 44 1904 431402|
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Manufacturer: Cytoskeleton Inc
Cardiac myosin protein has been purified from bovine heart tissue(1, 2). The full length myosin protein was purified with its essential light chains (ELC) and regulatory light chains (RLC), see Figure 1and 2. Myosin was then digested with alpha-chymotrypsin to liberate the soluble subfragment-1 (S1) domain, which was isolated by centrifugation (3). The purified myosin S1 fragment has been determined to be biologically active in an F-actin activated ATPase assay (see biological activity assay). Bovine cardiac myosin S1 fragment protein is supplied as a white lyophilized powder.
Storage and Resonstitution
Briefly centrifuge to collect the product at the bottom of the tube. Reconstituting a 1 mg tube of MYS03 with 300 ul of Milli-Q water (3 ml for the 10 mg bottles) will generate a 3.3 mg/ml stock of cardiac S1 myosin in the following buffer: 20 mM PIPES pH 7.0,5% (w/v) sucrose and 1% (w/v) dextran. The protein should not be exposed to repeated freeze-thaw cycles. The lyophilized protein is stable at 4°C desiccated (<10% humidity) for 1 year.
Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% gradient polyacrylamide gel. The myosin and its light chains used to produce the myosin S1 fragment was determined to be 90% pure (see Figure2).
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