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You are here:Home » Kits and Assays » Kits for Antibody Sets, BioAssay™ » ZO Tight Junction Antibody Kit, BioAssay™

ZO Tight Junction Antibody Kit, BioAssay™

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Specifications

Tight junctions are specialized regions of cell-cell contact that seal the paracellular space. TJ strands on adjacent cells are presumed to interact with each other to form a "molecular gasket" that prevents ions, water and other molecules from leaking between cells. In addition to this barrier function, the fence function of tight junctions plays an important role in maintaining epithelial cell polarity by blocking the diffusion of membrane proteins between apical (luminal) and basolateral cell surfaces. At one time, the only transmembrane protein known to be associated with tight junctions was occludin, a ~65kD protein with four transmembrane domains. Despite widespread expectation, a critical structural role for occludin in TJ strands was ruled out by the observation of apparently normal tight junctions formed between cells disrupted at both occludin alleles. Fortunately, a closer examination of isolated tight junctions uncovered two related ~22kD, four-transmembrane domain proteins, claudin-1 and claudin-2, with no similarity to occludin. In contrast to occludin, which induces only a small number of short strands at cell-cell contact sites when introduced into fibroblasts lacking tight junctions, claudin-1 and -2 induce networks of strands characteristic of true tight junctions.1,2 These findings suggest that
claudin-1 and -2 are major structural components of TJ strands and that occludin plays an accessory role. Claudins interact directly with the membrane-associated guanylate kinase homologues, ZO-1, ZO-2, and ZO-3, and are complexed with two other transmembrane proteins, occludin and junctional adhesion molecule.3 E-Cadherin is also important for the formation and maintenance of tight junctions.4 Cell-cell contacts containing E-Cadherin are crucial to the proper assembly and permanent sealing of tight junctions.5 To date, 20 claudins have been identified; studies suggest that the claudins comprising tight junctions are differentially expressed in different tissues. A number of human diseases are caused by claudin defects, which underscores the importance of claudins in human physiology. Deletion of the claudin-11 gene in mice results in disappearance of tight junction fibrils in testis and male sterility, as well as delayed axonal conduction rates in the CNS.7 An autosomal recessive form of deafness has been associated with human mutations in claudin-14.8 Claudin-7 expression is reportedly lower in invasive ductal breast carcinomas.
Catalog #Z0775
Kit ComponentsZ0775A: Ms/Mono; Z0775B: Rb/Poly; Z0775C: Ms/Mono; Z0775D: Rb/Poly; Z0775E: Rb/Poly; Z0775F: Ms/Mono.
All Mabs are IgG1, kappa and 20ug. The Pabs are IgG and 10ug.
Z0775A, Z0775B ZO-1; Z0775C, Z0775D: Occludin; Z0775E: Claudin-1; Z0775F: E-Cadherin
Z0775A Human, canine
Z0775B: Human, mouse, rat, canine
Z0775C: Human, rat, canine
Z0775D: Human, canine
Z0775E: Human, rat, canine
Z0775F: Human
ApplicationsZ0775A: Ms/Mono;Z0775C: Ms/Mono; Z0775F: Ms/Mono.
Suitable for use in ELISA, Immunofluorescence and Western Blot
Z0775B Rb/Poly; Z0775D: Rb/Poly; Z0775E: Rb/Poly;Suitable for use in ELISA, Immunofluorescence, Immunoprecipitation, Western Blot and Immunohistochemistry.
Recommended DilutionELISA: 0.1-1ug/ml
Immunofluorescence: 10-20ug/ml
Western blot: 0.5-3ug/ml
Immunohistochemistry (with ZO-1 and Occludin pAbs and E-Cadherin mAb): 2-5ug/ml
Immunoprecipitation (with ZO-1 and Occludin pAbs and E-Cadherin mAb): 2-5ug/ IP reaction.
Optimal dilution determined by the researcher.
Pretreatment protocols for use with formalin-fixed, paraffin-embedded tissue sections. 1. ZO-1 and Occludin pAbs Protease (Sigma P-5147) at 1-2mg/ml for 10 minutes at 37°C, after slides have been deparaffinized and rehydrated in ethanol. Normal human colon sections may be used as a positive control.
2. E-Cadherin mAb: HIER with citrate buffer, pH 6.0 for 15 minutes at 95°C, or boiling. Normal human colon sections may be used a positive control.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
PurityAll antibodies are purified by affinity chromatography. The monoclonal antibodies are Protein A purified from mouse ascites and the polyclonal antibodies are antigen affinity purified.
Concentration~0.5mg/ml
FormAll antibodies are supplied as a liquid in PBS, pH 7.4, 0.1% sodium azide.
SpecificityZO-1, ZO-1, Occludin, Occludin, Claudin-1, E-Cadherin. Reactivity: Human, rat, canine, mouse.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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