Catalogue

Background
Cadherins constitute a family of transmembrane glycoproteins involved in Ca2+-dependent cell-cell interactions. The members of this family are differentially expressed in various tissues. They function in the maintenance of tissue integrity and morphogenesis. Cadherins are divided into type I and type II subgroups. Type I cadherins include epithelial cadherin (E-cadherin, cadherin-1 or uvomorulin), neural cadherin (N-cadherin or cadherin-2), placental cadherin (P-cadherin or cadherin-3) and retinal cadherin (R-cadherin or cadherin-4), whereas kidney cadherin (K-cadherin or cadherin-6) and osteoblast cadherin (OB-cadherin or cadherin-11) are type II cadherins. One of the best characterized cadherins is E-cadherin, a 120 kD transmembrane glycoprotein consisting of an 80 kD extracellular and a 40 kD transmembrane and cytoplasmic part. The extracellular domains of E-cadherin are responsible for calcium binding which allows for homophilic interaction with other E-cadherin molecules on the same cell and neighbouring cells. In addition, E-cadherin can interact heterophilically with integrin αEβ7. The cytoplasmic domain of E-cadherin is linked to the actin cytoskeleton through the associated cytoplasmic Catenin proteins, thus establishing a complex localized to adherens junctions. In carcinomas E-cadherin is frequently downregulated, which is consistent with its function of an invasion suppressor in normal epithelia.

Source
5H9 is a Mouse monoclonal IgG1 antibody obtained by fusion of P3-X63-Ag 8,653 Mouse myeloma cells with spleen cells from a BALB/c Mouse immunized with affinity purified 80 kD extracellular fragments of E-cadherin derived from tryptic digestion of A-431 Human vulva carcinoma cells.

Product
Each vial contains 100 ul of 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.

Formulation: Each vial contains 100 ul of 1 mg/ml purified monoclonal antibody in PBS containing 0.09% sodium azide.

Specificity
5H9 recognizes both the 120 kD E-cadherin and its 80 kD trypsin-resistant extracellular part.

Applications
5H9 is suitable for immunoblotting, immunocytochemistry and immunohistochemistry on frozen and paraffin embedded tissues. For frozen tissues we recommend fixation with 1% formaldehyde in PBS buffer containing 0.1 mM CaCl2 and 0.1 mM MgCl2 for 10 minutes at room temperature, followed by a permeabilization step with 0.2% Triton X-100 in the same buffer for 5 minutes . For paraffin embedded tissues use a pretreatment step of 15 minutes incubation in TRIS-EDTA buffer pH 9 in a microwave. Optimal antibody dilution should be determined by titration, recommended range is 1:50 – 1:100 for immunohistochemistry with avidin-biotinylated Horseradish peroxidase complex (ABC) as detection reagent, and 1:100 – 1:500 for immunoblotting applications.

Storage
The antibody is shipped at ambient temperature and may be stored at +4°C. For prolonged storage prepare appropriate aliquots and store at or below -20°C. Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance or the concentration of the product.

Caution
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but our company accepts no liability for any inaccuracies or omissions in this information.

References
1. Frixen, U. H., Behrens, J., Sachs, M., Eberle, G., Voss, B., Warda, A., Lochner, D., and Birchmeier, W. (1991). E-cadherin-mediated cell-cell adhesion prevents invasiveness of Human carcinoma cells, J Cell Biol 113, 173-85. 2. Moll, R., Mitze, M., Frixen, U. H., and Birchmeier, W. (1993). Differential loss of E-cadherin expression in infiltrating ductal and lobular breast carcinomas, Am J Pathol 143, 1731-42. 3. Gabbert, H. E., Mueller, W., Schneiders, A., Meier, S., Moll, R., Birchmeier, W., and Hommel, G. (1996). Prognostic value of E-cadherin expression in 413 gastric carcinomas. Int J Cancer 69, 184-189. 4. Schwechheimer, K., Zhou, L., and Birchmeier, W. (1998). E-Cadherin in Human brain tumours: loss of immunoreactivity in malignant meningiomas. Virchows Arch 432, 163-167.

Protein Reference(s)

Database Name: UniProt

Accession Number: P12830

Safety Datasheet(s) for this product: