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Western Blotting and Antibodies

BM Benjamin B. Morris
NW Nolan A. Wages
PG Patrick A. Grant
PS P. Todd Stukenberg
RG Ryan D. Gentzler
RH Richard D. Hall
WA Wallace L. Akerley
TV Thomas K. Varghese
SA Susanne M. Arnold
TW Terence M. Williams
VC Vincenzo Coppola
DJ David R. Jones
DA David T. Auble
MM Marty W. Mayo
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Cell extracts were isolated using RIPA lysis method and westerns were performed as described previously (34). Primary antibodies were used at 1:1000 according to manufacturer specifications. Primary antibodies include: MYBL2 (Millipore #MABE886), CHK1 (Novus Biologicals #NB100-464), αTubulin (Sigma-Aldrich #T9026), γH2AX pS139 (Cell Signaling #9718), and H2AX (Cell Signaling #7631). Secondary antibodies conjugated to HRP include anti-rabbit IgG (Cell Signaling #7074) and anti-mouse IgG (Cell Signaling #7076). Secondary antibodies were used at 1:2500. Densitometric analyses were performed on autoradiographs and fold change relative to tubulin loading control was calculated using NIH ImageJ 1.46r software.

Copyright and License information:  ©2021 Morris, Wages, Grant, Stukenberg, Gentzler, Hall, Akerley, Varghese, Arnold, Williams, Coppola, Jones, Auble and Mayo
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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