I am simply trying to inhibit Src signaling to assess structural formation in SUM159 cells. A colleague noted that it may not be good for IF staining since the fairly dark orange coloration of the chemical might/would interfere with imaging on our IF scope. I might be able to use our Confocal and and adjust the laser output if necessary, but I was wondering if anyone had used SU6656, at what concentrations and for how long, and if anybody had any issues using this inhibitor with IF.
Thanks for the input.
Hi Kyle,
I can suggest using Src inhibitor PP2. In contrast to SU6656 its solution is colorless and therefore should not interfere with your IF. Furthermore, specificity of the inhibitor is quite good. Sigma states that it is 10,000-fold more specific for Src than for ZAP-70 or Jak2.
I myself have made good experiences using this inhibitor at concentrations ranging between 0.5 and 3 µM, incubation for 45 min at 37°C prior to analysis.
Hope this helps.
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