primary antibodies against pstat1  (Cell Signaling Technology Inc)

Journal: British Journal of Cancer

Article Title: Impaired T-bet-pSTAT1 α and perforin-mediated immune responses in the tumoral region of lung adenocarcinoma

doi: 10.1038/bjc.2015.255

Figure Lengend Snippet: Reduced STAT1 phosphorylation in the tumoral region of NSCLC. ( A – F ) Western blot analysis of pSTAT1 and actin expression in lung tissue samples from the tumoral, peri-tumoral and control region of patients with adenocarcinoma (ADC) ( N Control =5; N Peri-tumoral =5, N Tumoral =5). Bar charts show mean values of the protein expression levels of pSTAT1 α -Isoform ( C and D ) and pSTAT1 β -Isoform ( E and F ) relative to actin levels in ADC and in SCC, respectively. ( G ) Flow cytometry analysis of pSTAT1 + cells in the tumoral, the peri-tumoral and the control lung region of one representative patient with NSCLC. pSTAT1 staining was performed with the lung cell suspensions after 20 min of incubation at 37 °C in the presence or absence of IFN- γ . pSTAT1 + cells were gated on CD4 + , CD8 + or CD11b + cells, respectively. ( H ) Flow cytometry analysis of pSTAT1 + CD4 + and pSTAT1 + CD8 + T cells gated on lymphocytes as well as pSTAT1 + CD11b + cells gated on big cells in the control, the peri-tumoral and the tumoral lung region of one representative patient with NSCLC. Data are shown as mean values±s.e.m. using Student's t -test * P =0.05.

Article Snippet: Polyacrylamide gel electrophoresis was performed at 80 V for 1.5 h. Proteins were then transferred to a nitrocellulose membrane (PAGEr Ex Gels, Lonza, Rockland, USA) at 200 mA for 45 min. After a washing step and blocking of the membrane for 1 h (5% milk powder, 0.1% Tween-20 in PBS) at room temperature, the primary antibody against pSTAT1 (Cell Signaling Technology, Danvers, MA< USA) was applied and incubated overnight at 4 °C.

Techniques: Phospho-proteomics, Western Blot, Expressing, Control, Flow Cytometry, Staining, Incubation