Schedl Lab

Washington University Genetics

Activated MPK-1 Staining (MAPK-YT, Diphospho-MPK-1)

Prepared by Sudhir Nayak

The following modification to the Schedl Lab dissection and staining protocol applies to Sigma monoclonal M8159 for detecting activated MAP kinase.

Staining for activated MPK-1 with Sigma monoclonal anti-activated MAP kinase (diphosphorylated ERK-1&2) antibody (M8159) in C. elegans is particularly sensitive to dissection time.  Dissection times after the addition of levamisole must be limited to 5 min or less.  Dissection times of longer than 5 min will result in reduced signal.

After dissection follow the formaldehyde fixation and staining protocol.  Use the Sigma monoclonal anti-activated MAP kinase antibody from 1:100 to 1:400 depending on the sensitivity of your detecting equipment. Formaldehyde/methanol (mixed fixation) protocol can also be used if required but nuclear morphology is not as crisp and anti-activated MAP kinase signal is lower than with formaldehyde alone.

  • Tim Schedl, Ph.D
    Department of Genetics
    Campus Box #8232
    Washington University School of Medicine
    4566 Scott Ave.
    St. Louis, MO 63110

    Contact

    Email: ts@genetics.wustl.edu
    PHONE: (314)362-6164 [lab]
    FAX: (314)362-7855