WB (peptide competition) analysis of Jurkat cells treated with 10 mM H2O2 for 3 minutes (Lane 2-6) using GTX12862 Syk (phospho Tyr323) antibody prior incubated with the non-phosphopeptide corresponding to the phosphopeptide immunogen (Lane 3), a generic phosphotyrosine-containing peptide (Lane 4), or the phosphopeptide immunogen (Lane 5) control. The data show that only the immunogen phosphopeptide blocks the signal, demonstrating the specificity of the antibody. The membrane treated with lambda phosphatase (Lane 6) eliminates the signal further verifying that the antibody is phospho-specific.

ICC/IF analysis of Jurkat cells treated with 100 uM H2O2 for 1 hour using GTX12862 Syk (phospho Tyr323) antibody. Panel e shows untreated cells with no signal. Panel f represents control cells with no primary antibody to assess background.
Green : Primary antibody
Blue : Nuclei
Red : Actin
Fixation : 4% paraformaldehyde
Permeabilization : 0.1% Trito X-100 for 10 minutesDilution: 2 μg/ml