Tight Junction Protein ZO-1 ELISA Kit (TJP1)

INTENDED USE

Human TJP1 ELISA kit can measure concentrations of TJP1 (Tight Junction Protein ZO-1, ZO1, Zona occludens protein 1) present is plasma, serum, biological fluids or tissue homogenate samples. This assay has a minimum sensitivity detection limit of 0.094 ng/ml.

INTRODUCTION

TJP1, known as tight junction protein ZO-1, is a key scaffolding component of specialized intercellular complexes called tight junctions. By linking the cytoskeletons of adjacent epithelial or endothelial cells, it helps form regulated tissue barriers vital for physiological separation of fluid compartments throughout the body.

Located at the uppermost region of intercellular space, tight junctions create a selective gate between cells, allowing absorption and secretion while blocking free transit of solutes across epithelial sheets. TJP1 connects tight junction transmembrane proteins like claudins and occludin to the intracellular actin network. This orients and supports transmembrane components while transducing regulatory signals bidirectionally across the cell membrane.

Through cytoskeletal tethering and protein interactions, TJP1 organizes transmembrane tight junction proteins and reinforces cell polarity integral for barrier function. However, it also dynamically communicates signals reporting extracellular conditions to intracellular pathways managing epithelial permeability. For example, reflecting immune responses to intestinal pathogens, inflammatory signals trigger ZO-1 phosphorylation, causing it to loosen its cytoskeletal ties and relax the tight junction gate. This temporary leakiness enables immune cell transit toward infection sites.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti-TJP1.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this TJP1 ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.156, 0.312, 0.625, 1.25, 2.5, 5, 10 ng/ml.
Reactivity: Human
Sensitivity: 0.094 ng/ml
Range: 0.156 – 10 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for TJP1, no cross reactivity or interference between TJP1 and analogues was detected.
– Recovery: Serum (85 – 99%), EDTA Plasma (89 – 104%), Heparin Plasma (88 – 97%).
– Linearity: Serum (91 – 100%), EDTA Plasma (92 – 102%), Heparin Plasma (87 – 101%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. Tight junction proteins occludin and ZO-1 as regulators of epithelial proliferation and survival. Ann N Y Acad Sci. (2022) 1514 (1): 21-33. Kuo W.T., et al.
2. The tight junction protein TJP1 regulates the feeding-modulated hepatic circadian clock. Nat Commun. (2020) 11 (1): 589. Liu Y., et al.
3. ZO-1 Guides Tight Junction Assembly and Epithelial Morphogenesis via Cytoskeletal Tension-Dependent and -Independent Functions. Cells. (2022) 11 (23): 3775. Haas A.J., et al.
4. Estrogen decreases tight junction protein ZO-1 expression in human primary gut tissues. Clin Immunol. (2017) 183: 174-180. Zhou Z., et al.

ADDITIONAL INFORMATION

• Full Name: Tight Junction Protein ZO-1 ELISA Kit (TJP1)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 0.094 ng/ml

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