|Tel :||01904 431402||Bioquote Limited|
The Raylor Centre
York, YO10 3DW
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|Int Tel :||00 44 1904 431402|
|Int Fax :||00 44 1904 431409|
Manufacturer: Cytoskeleton Inc
* Limited stock available. If stock is not available, Cytoskeleton will produce a new batch upon request. Minimum order will apply. Inquire for more information.
Product Uses :
Cytoplasmic dynein was purified from porcine brain by the procedure of B.M. Paschal et al. (1). The preparation contains dynein heavy chain (520 kDa), intermediate chain (54, 59, and 74 kDa) and light chains (c.20 kDa). The protein has been determined to be biologically active in a microtubule-activated ATPase assay (see below). The protein is supplied as a lyophilized white powder.
Storage and Reconstitution :
The protein is supplied in 50 µg amounts which are designed for single use experiments. The powder is resuspended on ice with 50 µl of ice cold water to create a 1 mg/ml solution of dynein with a final buffer composition of 20 mM Tris-HCl pH 7.5, 50 mM KCl, 2 mM MgCl2, 0.5 mM EGTA plus 20% (w/v) sucrose. In this formulation the dynein will remain active for 3 days when stored at 4°C. Do not refreeze the protein as this will irreversibly denature the protein.
Protein purity is determined by scanning densitometry of a Coomassie-stained SDS-PAGE gradient gel. Figure 1, lane 9 shows 10 mg of CS-DN01 protein and purity was determined to be c.80%. The total protein in each tube will therefore be approximately 20% greater than the amount shown on the tube. Contaminants include kinesin, dynamin, dynactin and tubulin.
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