Amylase antibody

Cerulein was purchased from MedChem Express (HY-A0190, Shanghai). Lipopolysaccharide (LPS) was purchased from Sigma (L2630, Shanghai). ABT702 was purchased from Merck Millipore (116890, Shanghai). ZM241385 (an adenosine A_2A receptor antagonist) and GSK2606414 (PERK inhibitor) were purchased from Selleck (S8105 and S7307, respectively, Shanghai). The following primary antibodies were purchased: ß-actin antibody (Proteintech, 66009), ADK antibody (Abcam, ab227087), RIP1 antibody (CST, 3,493), phosphor-RIP1 (Ser161) antibody (Affinity, AF7377), RIP3 antibody (CST, 15828), phosphor-RIP3 antibody (Abcam, ab195117), mouse MLKL antibody (Proteintech, 66675), rat MLKL antibody (Abcam, ab243142), phosphor-MLKL antibody (Affinity, AF7420), NF-κB p65 antibody (CST, 8242), phosphor-NF-κB p65 antibody (CST, 3,033), GRP78 Antibody (Proteintech, 11587), CHOP antibody (Proteintech, 15204), PERK antibody (Proteintech, 24390), phosphor-PERK antibody (CST, 3,179), eIF2α antibody (CST, 5324), phosphor-eIF2α antibody (CST, 3,398), CD11b antibody (Abcam, ab133357), MOMA-2 antibody (Abcam, ab33451), and amylase antibody (Santa Cruz Biotechnology, sc-46657). Secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, United States).

Cell necroptosis in the pancreas was detected using double staining with Evans blue dye (EBD) and amylase antibody (Santa Cruz Biotechnology, sc-46657). EBD (10 mg/ml) was injected intraperitoneally for 4 h before the mice were euthanized. After the operation, the pancreas was separated and frozen in liquid nitrogen, and then cryosections of the pancreas were immunostained with an amylase antibody. Sections were observed and imaged by a fluorescence microscope (Olympus) and analyzed using ImageJ software (National Institutes of Health, Bethesda, MD, United States).