Anti rabbit secondary antibody 7074

Cells were lysed in sample buffer containing 60 mM Tri-HCl, pH 6.8, 2% SDS, and 5% glycerol, and boiled for 10 min. Total protein concentration was measured using a BCA kit (P0010; Beyotime, Shanghai, China). Equal amount of protein from each sample was loaded and analyzed by Western blot as previously described [22 (link)]. Primary antibodies against p-STAT3 (9145), STAT3 (12640), p-JNK1/2 (9251), JNK1/2 (9252), p-p38 (9211), p38 (9212), p-ERK1/2 (9101), ERK1/2 (9102), inducible nitric oxide synthase (iNOS; 2977) and β-actin (4970), and anti-rabbit secondary antibody (7074) were all purchased from Cell Signaling (Boston, MA, USA). The West Dura Extended Duration substrate detection kit was from Thermo (34076; Rockford, IL, USA).

Polyclonal antibodies to 3-MST (ab85377) and CSE (ab151769) were from Abcam and used at a concentration of 1:1,000. Anti-rabbit secondary antibody (7074) was used at a concentration of 1:10,000 in TBS-T and was from Cell Signaling Technology.

Protein samples were analyzed as previously described (18 (link)). Antibodies against phosphorylated p38 (4511), p38 (8690), phosphorylated ERK1/2 (4370), ERK1/2 (4695), phosphorylated JNK (4668), JNK (9258), phosphorylated FAK (Tyr397, 8556), FAK (3285), IκB (4814), and GAPDH (8884), anti-mouse secondary antibody (7076) and anti-rabbit secondary antibody (7074) were purchased from Cell Signaling Technology (Danvers, MA). Antibody against TF (sc-393657) was purchased from Santa Cruz. Images were taken using Amersham Imager 600.