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. Author manuscript; available in PMC: 2013 Aug 27.
Published in final edited form as: Nitric Oxide. 2011 Nov 10;26(1):20–26. doi: 10.1016/j.niox.2011.11.001

Figure 1.

Figure 1

Detection of PTP1B nitrosylation. COS 7 cells were treated with oxidants (200 uM CSNO, 1 mM H2O2 or 1 mM diamide) as indicated for 15 min at 37 °C. Cell extracts were prepared in RIPA buffer containing NEM, reduced with either PE PEG or DTT, and then labeled with MPB. A and B: MPB-labeled (oxidized) PTP1B was shifted by avidin as described in the methods. The total (without avidin added) and reduced PTP1B (with avidin added, which shifted oxidized PTP1B away) were visualized by immunoblotting with PTP1B antibody. C: PTP1B was immunoprecipitated from treated extracts using a specific monoclonal antibody. MPB-labeled (oxidized) PTP1B was visualized by avidin-HRP (upper blot).