PhosphoGSK3beta Ser9, rabbit mAb, Cell Signaling #9322s

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2 User Reviews

  • This antibody was tested on 50 ug lysate from mouse primary hippocampal neurons.
    A clear and intense signal was observed at the correct size after a very short exposure time. I used a concentration of 1:1000. The buffer used to lyse the cells is shown below:

    • 20 mM MOPS, pH 7.0 (any other buffer at this pH could be substituted);
    • 2 mM EGTA (to bind calcium);
    • 5 mM EDTA (to bind magnesium and manganese);
    • 30 mM sodium fluoride (to inhibit protein-serine phosphatases);
    • 60 mM β-glycerophosphate, pH 7.2 (to inhibit protein-serine phosphatases);
    • 20 mM sodium pyrophosphate (to inhibit protein-serine phosphatases);
    • 1 mM sodium orthovanadate (to inhibit protein-tyrosine phosphatases);
    • 1% Triton X-100 (can be substituted with 1% Nonidet P-40) Important Note: Do not add if you intend to first prepare a cytosolic fraction.
    • 1 mM phenylmethylsulfonylfluoride (to inhibit proteases);
    • 3 mM benzamidine (to inhibit proteases);
    • 5 µM pepstatin A (to inhibit proteases);
    • 10 µM leupeptin (to inhibit proteases);
    • 1 mM dithiothreitol (to reduce disulphide linkages)

    adjust to pH 7.2

    Western blot55555
    ImmunostainingNot Rated
    IPNot Rated
    ELISANot Rated
    Flow cytometryNot Rated
    LuminexNot Rated
  • I have used anti-phosphoGK3beta Ser9 (CST #9322s) at 1:1000 (milk based buffer) on primary murine macrophages. I get a nice band at 45 kDa as expected.

    Western blot44444
    ImmunostainingNot Rated
    IPNot Rated
    ELISANot Rated
    Flow cytometryNot Rated
    LuminexNot Rated

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Western blot
Immunostaining
IP
ELISA
Flow cytometry
Luminex