Horseradish peroxidase hrp conjugated goat anti rabbit igg h l

Poly (ethylene glycol)-block-poly (d, l)-lactic-co-glycolic acid (mPEG-b-PLGA, PEG 2000 Da, PLGA 8000 Da, LA/GA = 75/25) was provided by the laboratory of Professor Fangming Zhu at the School of Chemistry, Sun Yat-sen University, and tacrolimus was obtained from Selleck Chemicals (Shanghai, China). Dialysis bags were purchased from Spectrapor (MWCO 14000, Waltham, MA, USA). Isoflurane inhaled anesthesia (Yipin pharmaceutical Co., LTD, Hebei, China) and 3% sodium pentobarbital solution (Merck, Darmstadt, German) were used to anesthesia animals. Proparacaine hydrochloride eye drops (S.A. ALCON, Fort Worth, TX, USA), Iviz (Bausch Lomb, Shandong, China), and 10-0 black mono nylon sutures (Alcon, Fort Worth, TX, USA) were used in corneal transplantation of rats. Tobramycin eye ointment purchased from Zhongshan Ophthalmic Center (Guangzhou, China). 0.1% tacrolimus eye drops were purchased from Senju Pharmaceutical Co., Ltd. (Osaka, Japan). Rabbit polyclonal anti-CD4/CD8 antibody and horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG H&L were purchased from Abcam (Cambridge, MA, USA), the Milliplex^® xMAP Rat Cytokine/Chemokine kit was purchased from EMD Millipore Corporation (Billerica, MA, USA).

Western blotting was performed to confirm the results of quantitative proteomics. Briefly, protein samples of CHO cells and Caco2 cells stimulated with E. histolytica or Igl were separated on 10% polyacrylamide gels and then electrotransferred to polyvinylidene difluoride membranes (General Electric Co., Schenectady, NY, USA). After blocking with bovine serum albumin (5% in PBS), membranes were incubated with the following primary antibodies: anti-β-actin antibody (Abcam, Ab8227; Cambridge, United Kingdom), anti-PKM2 monoclonal antibody (Cell Signaling Technology, catalog no. 4053; Boston, MA, USA), and anti-LC3A/B antibody (Cell Signaling Technology, catalog no. 12741; Boston, MA, USA). Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG(H+L) (Abcam, Ab6721; Cambridge, United Kingdom) was used as the secondary antibody. Proteins were detected using an enhanced chemiluminescence (ECL) Western blotting substrate kit (Tanon, Shanghai, China).