Occludin (OCLN), human : Turner Lab

Occludin is an integral membrane protein that is required for cytokine-induced regulation of the tight junction paracellular permeability barrier. Mutations in the gene (OCLN) are a cause of band-like calcification with simplified gyration and polymicrogyria (BLC-PMG), an autosomal recessive neurologic disorder that is also known as pseudo-TORCH syndrome. Alternative splicing results in multiple transcript variants. A related pseudogene is present 1.5 Mb downstream on the q arm of chromosome 5.

Antibody validation
This is a rat antibody raised against the human occludin cytoplasmic tail. Our validation included staining of formalin-fixed, paraffin-embedded human duodenum, where the stain colocalized with other tight junction proteins, of control and inflammatory bowel disease human colon biopsies documenting downregulation in disease, and failure to label occludin knockdown Caco-2 cell lines. The antibody reacts with human and dog, but not mouse, occludin.

Immunostaining procedure conditions:
Antigen retrieval with Target Retrieval Solution, pH9 (Agilent Technologies, Cat. #S2367) for 40 minutes at 95°C. Primary and secondary incubations performed using the following reagents:

Antibody	Source	Catalog #	Lot #	RRID #	Working Dilution/Concentration
Anti-occludin	Turner lab	3H5D11		AB_2783881	1.5 µg/ml
Anti-E Cadherin antibody [M168]	Abcam	ab76055	GR216035	AB_1310159	0.83 µg/ml
Anti-Actin antibody	Abcam	ab8227		AB_867488	8.2 µg/ml


Alexa Fluor 488 AffiniPure Donkey Anti-Mouse IgG (H+L)	Jackson Immunoresearch	715-545-151	118237	AB_2341099	1.5 µg/ml
Alexa Fluor 594 AffiniPure F(ab’)₂ Fragment Donkey Anti-Rat IgG (H+L)	Jackson Immunoresearch	712-586-153		AB_2340622	1.5 µg/ml
Alexa Fluor 647 AffiniPure F(ab’)₂ Fragment Donkey Anti-Rabbit IgG (H+L)	Jackson Immunoresearch	711-606-1532		AB_2340696	1.5 µg/ml
Hoechst 33342	Life Technologies	H3570			10 µg/ml