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Determination of Syk phosphorylation by immunobloting
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Phosphorylation of Syk in the 25 μg cytoplasmic fraction in NP1 lysis buffer (Cell Signaling Technology) was analyzed by Western blotting using monoclonal phospho-Syk (Tyr525/526) (C87C1), or polyclonal phospho-Zap-70 (Tyr319)/Syk (Tyr352) Ab and Syk Ab, all from Cell Signaling Technology. In some experiments, the whole cell lysate was immunoprecipitated with phospho-Tyr mouse mAb (P-Tyr-100, Cell Signaling Technology) according to the manufacturer´s instructions and the immunoprecipitate was analyzed by Western blotting using Syk Ab.
Copyright and License information: ©2016 Aouar et al
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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