Product Uses
- Measurement of microtubule-activated ATPase/GTPase assays
- Identification/characterization of proteins or small molecules that affect motor ATPase/GTPase activity
- Identification/characterization of proteins or small molecules that affect motor/microtubule interactions.
Material
The conserved motor domain of human kinesin KIF7 was expressedin E.coli. The recombinant protein contains a 6 histidine tag at the amino terminal end and has a combined molecular weight of 44 kDa. The protein has been determined to be biologically active in a microtubule-activated ATPase/GTPase activity test. The protein is supplied as a lyophilized powder.
Storage and Reconstitution
The lyophilized protein is stable for 1 year when stored at 4°C with a desiccant (humidity <10%). Alternatively, the lyophilized protein can be stored at -70°C and is stable for 1 year. The protein should be reconstituted to 5 mg/ml with distilled water or CMW Buffer 1(100 mM PIPES pH 7.0, 200 mM KCl, 2 mM MgCl2, 1 mM DTT, 20μM ATP). See the table above for resuspension volumes. The protein can be aliquoted into experiment sized aliquots, snap frozen in liquid nitrogen and stored at -70°C. Under these conditions the protein is stable for 4 months. For working concentrations the Kif7 protein should be diluted in CMW Buffer 1. NOTE: kinesins do not respond well to repeated freeze/thaws and for storage at -70°C the protein concentration should not be less than 5 mg/ml. Kinesin diluted below 5 mg/ml should not be re-frozen as it will lose its biological activity.
Purity
Protein purity is determined by scanning densitometry of a Coomassie-stained SDS-PAGE gradient gel. Figure 1 shows 20 ug of KF51protein and purity was determined to be >80%.The total protein in each tube will therefore be approximately 20% greater than the amount shown on the tube.
