Anti caspase 11 nb120 10454

Samples were separated by 12% SDS-PAGE, followed by electrophoretic transfer to polyvinylidene fluoride membranes and blocking and incubating membranes with primary antibodies. The following primary antibodies were used: anti-caspase-1 (AG-20B-0042; AdipoGen), anti-caspase-3 (9661 and 9662; CST), anti-caspase-11 (NB120-10454; Novus Biologicals), anti-AIM2 (13095; CST), anti-NLRP3 (AG-20B-0014; AdipoGen), anti-TFEB (A303-673A; Bethyl Laboratories), anti-LC3 (catalog NB600-1384; Novus Biologicals), and anti-GAPDH (5174; CST). HRP-labeled anti-rabbit or anti-mouse antibodies (Cell Signaling Technology) were used as secondary antibodies.

The AR42J cells were treated with caerulein for 8 h. The untreated cells were used as control. After intensive washing with PBS containing 2% (wt/vol) FBS, cells were stained with fluorochrome-conjugated antibodies [anti–caspase-1 (AG-20B-0042-C100) was from Adipogen; anti–caspase-11 (NB120-10454) was from Novus] for FACS analysis. All flow cytometry was performed on an FACScalibur flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA), and data were analyzed by FlowJo 7.6.1 software.

For immunoblot studies, antibodies against caspase-11 (C1354) and β-actin (A1978) were purchased from SIGMA. Antibody against caspase-1 (AG-20B-0042) was purchased from Adipogen. Antibodies against Gabarap (M135-3) and Gate-16 (18724-1-AP) were purchased from MBL or Proteintech. Antibody against GBP2 (sc-10588) was purchased from Santa Cruz. Antibody against ASC (04-147) was obtained from Millipore. TOM20 (11802-1-AP) was obtained from Proteintech. Antibody against β-COP (ab2899) were purchased from Abcam. Anti-Aim2 (ab93015) was purchased from Abcam. Anti-NLRC4 (06-1125) was purchased from Millipore.
For immunofluorescence studies, anti-GBP2 (11854-1-AP) were purchased from Proteintech. Anti-HA (MMS-101R) was purchased from Covance. Anti-caspase-11 (NB120-10454) was obtained from Novus. Anti-caspase-11 (C1354) and FITC-LPS (F3665) were obtained from Sigma. Anti-ASC (#67824) was obtained from Cell Signaling. Anti-ubiquitin mouse monoclonal antibody (FK2; MFK-004) was obtained from Nippon Biotest Labotatories. Anti-p62 (PM045) was obtained from MBL.