D lactate antibody

For immunoblotting, the intestinal tissue was lysed in radio immunoprecipitation assay lysis buffer. The lysate was separated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane. The protein concentration of lysate was detected using a bicinchoninic acid kit (Bio-Rad Laboratories, Hercules, CA, USA). The blot of lysates was incubated with the following antibodies: IL-9 antibody (Abcam, diluted in 1:1000), D-lactate antibody (abcam, diluted in 1:1000), zonula occludens 1(ZO-1) antibody (proteintech, diluted in 1:1000), horseradish peroxidase (HRP)-labeled anti-rabbit antibody (Beyotime, Shanghai, China), GAPDH antibody (ZEN-bio, diluted in 1:5000), and HRP-labeled anti-mouse antibody (Beyotime, Shanghai, China). Images were acquired with Tanon enhanced chemiluminescence substrate (Tanon, Shanghai, China), using Tanon 5200 imaging system (Tanon, Shanghai, China).

After rat sacrifice, the intestinal sample was disintegrated in radio immunoprecipitation assay lysis buffer. Following the lysate isolation in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the lysate was loaded onto a nitrocellulose membrane. Using a bicinchoninic acid kit (Bio-Rad Laboratories, Hercules, CA, USA), we determined the expression level of protein in the lysate sample. The lysate blotting was subjected to incubation using each antibody as follows: GAPDH antibody (ZEN-bio, diluted in 1:5000), HRP-labeled anti-mouse antibody (Beyotime, Shanghai, China), D-lactate antibody (Abcam, diluted in 1:1000), E-cadherin antibody (Abcam, diluted in 1:1000), αEβ7 antibody (affinity, diluted in 1:1000), IL-9 antibody (Abcam, diluted in 1:1000), and horseradish peroxidase (HRP)-labeled anti-rabbit antibody (Beyotime, Shanghai, China). Tanon-enhanced chemiluminescence substrate of the Tanon 5200 imaging system was used (Tanon, Shanghai, China) to produce the images.