New batches of this antibody are no longer guaranteed in ICC/IF, IHC-P or IHC-Fr as they have not passed our stringent batch testing criteria. Please contact customer support for any specific queries. We recommend ab216327 as an alternative for ICC/IF and IHC-P.
The immunogen used to raise this antibody has 78% homology with the Mouse Occludin protein and the Rat Occludin protein. Some customers have successfully used ab31721 with Mouse/Rat lysates/tissue, however we have not been successful detecting Occludin in these species in our own testing.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/250. Detects a band of approximately 62 kDa (predicted molecular weight: 59 kDa).
Use at an assay dependent concentration. PubMed: 22689949
May play a role in the formation and regulation of the tight junction (TJ) paracellular permeability barrier. It is able to induce adhesion when expressed in cells lacking tight junctions.
Localized at tight junctions of both epithelial and endothelial cells. Highly expressed in kidney. Not detected in testis.
Defects in OCLN are the cause of band-like calcification with simplified gyration and polymicrogyria (BLCPMG) [MIM:251290]; also known as pseudo-TORCH syndrome. BLCPMG is a neurologic disorder with characteristic clinical and neuroradiologic features that mimic intrauterine TORCH infection in the absence of evidence of infection. Affected individuals have congenital microcephaly, intracranial calcifications, and severe developmental delay.
Belongs to the ELL/occludin family. Contains 1 MARVEL domain.
The C-terminal is cytoplasmic and is important for interaction with ZO-1. Sufficient for the tight junction localization. Involved in the regulation of the permeability barrier function of the tight junction (By similarity). The first extracellular loop participates in an adhesive interaction.
Phosphorylated upon DNA damage, probably by ATM or ATR. Dephosphorylated by PTPRJ. The tyrosine phosphorylation on Tyr-398 and Tyr-402 reduces its ability to interact with TJP1.
All lanes : Anti-Occludin antibody (ab31721) at 1/250 dilution
Lane 1 : Wild-type HAP1 whole cell lysate at 40 µg Lane 2 : OCLN (Occludin) knockout HAP1 whole cell lysate at 40 µg Lane 3 : HeLa whole cell lysate (Low Occludin expression) at 20 µg Lane 4 : HepG2 whole cell lysate (High Occludin expression) at 20 µg
Predicted band size: 59 kDa
Lanes 1 - 4: Merged signal (red and green). Green - ab31721 observed at 59 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab31721 was shown to recognize Occludin in wild-type HAP1 cells as signal was lost at the expected MW in OCLN (Occludin) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and OCLN (Occludin) knockout samples were subjected to SDS-PAGE. Ab31721 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/250 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Occludin antibody (ab31721)
Lane 1 : Anti-Occludin antibody [EPR8208] (ab167161) at 1/5000 dilution Lane 2 : Anti-Occludin antibody (ab31721) at 1/1000 dilution Lane 3 : Anti-Occludin antibody [EPR20992] (ab216327) at 1/5000 dilution
All lanes : Recombinant Human Occludin protein (ab114189)
Lysates/proteins at 0.025 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Wu D et al. Formononetin Administration Ameliorates Dextran Sulfate Sodium-Induced Acute Colitis by Inhibiting NLRP3 Inflammasome Signaling Pathway. Mediators Inflamm2018:3048532 (2018).
Read more (PubMed: 29507526) »
Guo W et al. Analysis of differential expression of tight junction proteins in cultured oral epithelial cells altered by Porphyromonas gingivalis, Porphyromonas gingivalis lipopolysaccharide, and extracellular adenosine triphosphate. Int J Oral Sci10:e8 (2018).
Read more (PubMed: 29319048) »