| ZO-1

Tight junction protein ZO-1 also known as Zonula Occludens-1 (ZO-1), is a 220-kD peripheral membrane protein that is encoded by the TJP1 gene. It belongs to the family of zonula occludens proteins (ZO-1, ZO-2, and ZO-3), which are tight junction-associated proteins and of which, ZO-1 is the first to be cloned. It was first isolated in 1986 by Stevenson and Goodenough using a monoclonal antibody raised in rodent liver to recognise a 225-kD polypeptide in whole liver homogenates and in tight junction-enriched membrane fractions. It has a role as a scaffold protein which cross-links and anchors Tight Junction (TJ) strand proteins, which are fibril-like structures within the lipid bilayer, to the actin cytoskeleton.
 

| Typical data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, wash technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data are provided below for reference only.

 

Concentration (ng/mL)	10	5	2.5	1.25	0.625	0.312	0.156	0
OD	2.04	1.4	0.89	0.64	0.42	0.27	0.21	0.06
Corrected OD	1.98	1.34	0.83	0.58	0.36	0.21	0.15	-

 

| Repeatability

Intra-assay Precision (Precision within an assay)：CV% < 10%; Inter-assay Precision (Precision between assays)：CV% < 10%
 

| Recovery

The recovery of Human ZO-1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

 

Sample Type	Range(%)	Average Recovery(%)
Tissue Homogenates(n=8)	84-98	96
Cell Culture Supernates(n=8)	92-105	101

 

| Linearity

Samples were spiked with high concentrations of Human ZO-1 and diluted with Universal Diluent to produce samples with values within the range of the assay.

 

		Tissue Homogenates(n=4)	Cell Culture Supernates(n=4)
1 : 2	Range (%)	84-95	85-96
	Average (%)	91	93
1 : 4	Range (%)	89-103	87-108
	Average (%)	96	98