Mouse Anti-Cingulin Mouse Anti-Cingulin

Qty: 100 µg/200 µL Mouse anti-Cingulin Catalog No. 37-4300 Lot No. Mouse anti-Cingulin

FORM This monoclonal antibody is supplied as a 200 µL aliquot at a concentration of 0.5 mg/mL in PBS, pH 7.4, containing 0.1% sodium azide. This antibody is highly purified from mouse ascites by protein A chromatography.

CLONE: 3G1H5 ISOTYPE: IgG1-kappa/ IgG2a-kappa

IMMUNOGEN Recombinant protein encompassing the C-terminus of chicken Cingulin

SPECIFICITY This antibody is specific for the ~140 kDa Cingulin protein. On Western blots, it identifies a band at 140 kDa in human Caco- 2 cell lysates. Cross reactivity with related proteins has not been observed.

REACTIVITY Western blot utility has been confirmed with human Caco-2 cell lysates. This antibody also labeled Cingulin in the junctions of mouse renal epithelial cell samples. Based on amino acid sequence homology, this antibody is also expected to react with chicken, dog, rat, Xenopus, and zebrafish Cingulin.

Sample Western Immunofluorescence ELISA Blotting Human +++ ND ND Mouse ND +++ ND Dog ND ND ND Rat ND ND ND Chicken ND ND ND (Excellent +++, Good++, Poor +, No reactivity 0, Not applicable N/A, Not Determined ND)

USAGE Working concentrations for specific applications should be determined by the investigator. Appropriate concentrations will be affected by several factors, including secondary antibody affinity, antigen concentration, sensitivity of detection method, temperature and length of incubations, etc. The suitability of this antibody for applications other than those listed below has not been determined. The following concentration ranges are recommended starting points for this product.

Western Blotting: 1-3 µg/mL Immunofluorescence: 20-40 µg/mL

STORAGE Store at 2-8°C for up to one month. Store at –20°C for long-term storage. Avoid repeated freezing and thawing.

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www.invitrogen.com Invitrogen Corporation • 542 Flynn Rd • Camarillo • CA 93012 • Tel: 800.955.6288 • E-mail: [email protected] PI374300 (Rev 10/08) DCC-08-1089 Important Licensing Information - These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen Catalog or our website, www.invitrogen.com). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

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BACKGROUND

Cingulin is a protein component of vertebrate tight junctions, which are important in the maintenance of epithelial polarity and in the formation of barriers between different tissue compartments.1-2 Cingulin has been detected in the junctions and submembrane areas of human normal and neoplastic epithelia3 in the junctions of epithelial cells of the organ of Corti,3 chicken embryonic precursors of endothelial and cardiac cells,4 and urodele embryos.5 Cingulin has also been detected in the nuclei of cultured epithelial cells.6,7 Cingulin interacts in vitro with several junctional and cytoskeletal proteins, including ZO-1, ZO-2, ZO-3, AF-6, myosin, and actin.3,4 Cingulin interaction with ZO-1 may be important in recruiting cingulin into junctions.8

REFERENCES 1. Citi S, et al. Nature 333(6170): 272-6, 1988. 2. D'Atri F and Citi S. FEBS Lett. 507:21-24, 2001. 3. Eisenberg CA and Bader D. Dev Biol. 167:469-481, 1995. 4. Leonova EV and Raphael Y. Hear Res. 113:14-28, 1997. 5. Shook DR, et al. Dev Biol. 248:220-39, 2002. 6. Citi S and Cordenonsi M. 1999. In Adhesive Interactions of Cells. Vol. 28. Garrod DR, et al, editors. JAI Press, Inc., Greenwich, CT.203-233. 7. Nakamura T, et al., Proc Natl Acad Sci U S A. 97:7284-7289, 2000. 8. D’Atri F, et al. J Biol Chem 277(31): 27757-64, 2002.