Rho Kinase （ROCK） Activity Assays

Rho Kinase （ROCK） Activity Assays

Our ROCK Activity Assays use a non-radioactive

format to measure the level of active ROCK in cell

or tissue lysates. The Immunoblot kit provides a

convenient format for measuring ROCK activity in a

few samples, while the ELISA kit contains a strip-

well plate precoated with recombinant MYPT1 for

higher throughput.

Results Using the ROCK Activity Immunoblot Kit. Kinase

buffer containing 10 ng of active ROCK-II was incubated with 0.2

mM ATP and 500 ng of recombinant MYPT1 for 30 minutes at

30ºC. Reaction was stopped by adding 4X SDS-PAGE sample

buffer. Serial dilutions were loaded onto SDS-PAGE. Lanes 1, 3,

5, 7: Without ROCK （negative control）. Lanes 2, 4, 6, 8: With

ROCK. Lanes 1 & 2: 200 ng MYPT1; Lanes 3 & 4: 100 ng;

Lanes 5 & 6: 50 ng; Lanes 7 & 8: 25 ng. Phosphorylation of

MYPT1 substrate was detected by anti-phosphor-MYPT1 as de-

scribed in the assay protocol.

•  Safe: Non-radioactive assay format

•  Versatile: Measure ROCK activity in cell or tissue

lysates by Western blot or ELISA

•  Quantitative: Active ROCK included as positive

control

ROCK Activity Immnoblot Kit Assay Principle.