Your search for " proWeb " resulted in 3 protocols
ATPase Assays with 32P-ATP Author: Liz Greene and Steve Henikoff Source:proWeb Abstract: Prepare reaction mix. add motor last. vortex briefly.
Take timepoints at 0. 2. 5. 10. 20. 40. 60 min by withdrawing 40 µl into microfuge tubes containing 0.76 ml of the activated charcoal suspension.
Vortex tubes. keep on ice 15 min. centrifuge 5 min. withdraw supernatant into new tube and centrifuge again to remove charcoal particles.
Nucleotide Binding/Hydrolysis Assays Author: Liz Greene and Steve Henikoff Source:proWeb Abstract: Apply reaction mix to a Sephadex G-50 Medium column.
Collect two 1 mL fractions and label them fr 1 and fr 2. then collect 28 x 5-drop fractions (each = ~250 µL) and label them fr 3 - fr 30.
Steady State ATPase Assays Author: Liz Greene and Steve Henikoff Source:proWeb Abstract: Assemble microtubules for assays. E.g..58 µL of 50 µM MTs = 50 µL 5.8 mg/mL tubulin + 0.5 µL 100 mM Mg•GTP. incubate for 30-60 min at 37°C. then add 7.5 µL 544 µM Taxol in PM (1.02 µL 4 mM Taxol + 6.5 µL PM) at 37°C.
Determine protein concentration using the Bradford assay.
Top of Page
