IHC-FoFr: Use at an assay dependent dilution. PubMed: 20978075
WB
WB: 1/250. Detects a band of approximately 63 kDa (predicted molecular weight: 59 kDa).Can be blocked with Human Occludin peptide (ab34440). Abcam recommends using 3% milk as the blocking agent.
In-Cell ELISA
In-Cell ELISA: Use at an assay dependent concentration. PubMed: 22689949
ICC/IF image of ab31721 stained Hek293 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab31721, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Anti-Occludin antibody (ab31721)
Anti-Occludin antibody (ab31721) at 1 µg/ml (blocked with 3% milk) + Kidney (Human) Tissue Lysate - adult normal tissue (ab30203) at 20 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution developed using the ECL technique
Denis MC et al. Apple peel polyphenols and their beneficial actions on oxidative stress and inflammation. PLoS One8:e53725 (2013). WB ; Human . Read more (PubMed: 23372666) »
Chen JC et al. Coculturing human endometrial epithelial cells and stromal fibroblasts alters cell-specific gene expression and cytokine production. Fertil Steril100:1132-43 (2013). Read more (PubMed: 23849844) »
