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. Author manuscript; available in PMC: 2012 Sep 30.
Published in final edited form as: Cell. 2011 Sep 30;147(1):185–198. doi: 10.1016/j.cell.2011.08.036

Figure 5. scFv45 Displays Specificity for PTP1B-OX over TCPTP-OX.

Figure 5

(A) Recombinant TCPTP (1–317) was reversibly oxidized by H2O2 and reactivated by TCEP (5 mM). Phosphatase assay was determined using 100 nM 32P-RCML and 5 nM of enzyme from each sample.

(B) Purified scFvs (750 nM) were incubated with oxidized TCPTP (7.5 nM) and phosphatase assay was determined in presence of TCEP (5 mM). [Error bars in A and B show standard deviation from six (A) and three (B) phosphatase assays].

(C) TCPTP was reversibly oxidized with H2O2 and binding with scFv45 was assessed by anti-HA-Agarose pull down. Equivalent amounts (4 ng of TCPTP) of input, supernatant and precipitate were analyzed by immunoblot. As a control, a parallel pull down was performed with reversibly oxidized PTP1B and scFv45.

See also Figure S4.