Product Uses
- Measurement of Microtubule-activated ATPase assays
- Identification/characterization of proteins or small molecules that affect motor ATPase activity
- Identification/characterization of proteins or small molecules that affect motor/microtubule interactions.
Material
The conserved motor domain of human MKLP2 was expressed in a prokaryotic system. The recombinant protein contains a His-Tag at the amino terminal end and has a combined molecular weight of 54 kDa. The protein has been determined to be biologically active in a microtubule-activated ATPase activity test. The protein is supplied as a lyophilized powder.
Storage and Reconstitution
The lyophilized protein is stable for at least 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 at least 1 year. The protein should be reconstituted to 5 mg/ml with ice cold Milli-Q water. The protein can be aliquoted into experiment sized tubes and snap frozen in liquid nitrogen. When reconstituted and stored at -70°C, the protein will be stable for at least 4 months. 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 activity.
Purity
Protein purity is estimated by scanning densitometry of a coomassie-stained SDS-PAGE gradient gel. Figure 1 shows 5 μg of CS-MP05 protein and purity was determined to be >70%. The total protein in each tube will therefore be approximately 30% greater than the amount shown on the tube. The major contaminants are at approximately 51 & 70 kD. The microtubule-activated ATPase activity of the MKLP2 motor is not affected by these contaminants.
