Anti bax pab

Cultured cells or frozen rat intestinal tissue samples were lysed in radio-immunoprecipitation assay buffer, and protein was collected after centrifugation and mixed with 5 × sodium dodecyl sulfate (SDS) sample buffer. The samples were separated by SDS-polyacrylamide gel electrophoresis (PAGE) using 8–12% acrylamide gels and then transferred to polyvinylidene fluoride (PVDF) membranes (Millipore, Billerica, MA, USA). After incubation with primary and secondary antibodies, the protein bands were detected with chemiluminescence detection reagents (Millipore). The following antibodies (Abs) were used: goat anti-p22^phox, goat anti-gp91^phox pAb, and goat anti-p47^phox pAbs were all from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-PARP-1 pAb, anti-Bcl-2 pAb, anti-Bax pAb, anti-caspase-3 pAb, anti-JNK Ab, and anti-pJNK Ab were from Cell Signaling Technology (Beverly, MA, USA); anti-PAR mAb was from Millipore; rabbit anti-P47^phox pAb was from Sigma; and anti-AIF Ab was from Abcam (Cambridge, UK). Mouse anti-AOPP Ab was a gift from professor Fu Ning (Southern Medical University, Guangzhou, China). Mouse anti-β-actin Ab and goat anti-mouse, rabbit anti-goat, and goat anti-rabbit IgG-horseradish peroxidase (HRP) were purchased from Boster (Wuhan, China).

Astaxanthin and dimethyl sulfoxide were purchased from Sigma-Aldrich Inc. (Taufkichen, Germany). Cell Counting Kit-8 (CCK-8) was purchased from DOJINDO Inc. (Kumamoto, Japan). ROS Detection Assay Kit was purchased from Biovision Inc. (Milpitas, CA, USA). PE Annexin V apoptosis detection kit was purchased from BD Biosciences Inc. (San Jose, CA, USA). Anti-BAX pAb, anti-BCL2 mAb, anti-cleaved CASP3 mAb, and anti-cleaved PARP mAb were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Anti-ACTB pAb were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).