WB (peptide competition) analysis of NIH3T3 cells treated with 50ng/mL PDGF for 15 minutes (Lane 2-6) using GTX10564 CREB (phospho Ser129/Ser133) antibody prior incubated with the non-phosphopeptide corresponding to the phosphopeptide immunogen (Lane 3), a generic phosphoserine-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.

WB analysis of 20 μg of NIH/3T3 untreated (lane 1), treated for 10 minutes with 50 ng/ml of PDGF (lane 2), treated for 10 minutes with 200 ng/ml of EGF (lane 3), A549 untreated (lane 4), treated for 10 minutes with 200 ng/ml of EGF (lane 5), A431 (lane 6), SK-N-SH (lane 7), MCF7 (lane 8) and MDA-MB-231 (lane 9) cell lysate using GTX10564 CREB (phospho Ser129/Ser133) antibody.
Dilution : 1:1000

ChIP analysis of 2 million HeLa cells treated with 50ng/ml of TNF alpha for one hour using GTX10564 CREB (phospho Ser129/Ser133) antibody. Normal Rabbit IgG was used as a negative IP control. The precipitated DNA was detected by PCR with primer set targeting to the promoter of I-kB, YWHAZ gene as positive control target and the inactive GAPDH used as negative control target.

ICC/IF analysis of HeLa cells using GTX10564 CREB (phospho Ser129/Ser133) antibody. Panel e is a no primary antibody control.
Green : Primary antibody
Blue : Nuclei
Red : Actin
Fixation : 4% paraformaldehyde
Permeabilization : 0.25% Trito X-100 for 10 minutes
Dilution : 1:400 for 30 minutes at room temperature