Gapdh rabbit mab

The same batches of cells for functional studies were collected and resuspended in Lysis Buffer (150 mM NaCl, 20 mM HEPES, pH 7.25) in the presence of 5 mM MgCl₂, 0.1 mg mL⁻¹ DNase I, 1 × protease inhibitor cocktail. DDM was added to solubilize the membrane proteins at 4 °C for 2 h. The solubilization slurries were clarified by centrifugation, and the supernatant was subjected to SDS-PAGE followed by Western blot. We used the Mouse anti DDDDK-Tag mAb (ABclonal, AE005) to detect the expression of hAE2 and different mutants. The secondary antibody is Anti-mouse IgG, HRP-linked Antibody (Cell Signaling, #7076). We also use the GAPDH Rabbit mAb (Abclonal, Catalog number: A19056) to evaluate the cell quantity. The secondary antibody is Anti-rabbit IgG, HRP-linked Antibody (transgen, Catalog number: HS101-01). All the antibodies were diluted (1:10,000) with skim milk in TBST in prior to use. Images were captured with an Amersham Imager 600.

APL was extracted and purified from Antarctic krill oil, and its molecular species and composition were in agreement with previous reports [20 (link)]. Dextran sulfate sodium salt (DSS, MW: 40,000) was provided by Aladdin Biochemical Technology (Shanghai, China). Bicinchoninic acid (BCA) protein concentration assay kit was purchased from Beyonte Biotechnology (Shanghai, China). GAPDH Rabbit mAb, Occludin Rabbit pAb, and HRP Goat Anti-Rabbit IgG (H + L) were provided by ABclonal Technology (Wuhan, China). The ELISA kits of interleukin (IL)-6 and tumor necrosis factor (TNF)-α were supplied by Xinbosheng Biotechnology (Shenzhen, China). Urine fecal occult blood test kit were purchased from Nanjing Jiancheng Institute of Biological Engineering (Nanjing, China).

Specific amino acid sequences of CCV ORF39 (457–474 aa, and 1031–1045 aa) and ORF59 (299–316 aa and 75–90 aa) were initially synthesized and selected, then were coupled with KLH carrier protein. New Zealand white rabbits were vaccinated to collect rabbit serum to prepare rabbit anti-ORF39 and 59 polyclonal antibodies (ABclonal, Wuhan, China), as previously reported [16 (link)]. The following antibodies were purchased commercially: HRP Goat Anti-Mouse IgG (H + L) (ABclonal, China), Mouse anti His-Tag mAb (ABclonal, China), Mouse anti GFP-Tag mAb (ABclonal, China), GAPDH Rabbit mAb (ABclonal, China), HRP Goat Anti-Mouse IgG (H + L) (Beyotime, Haimen, China), and HRP Goat Anti-Rabbit IgG (H + L) (Beyotime, China).