Nectin-3 is a 83kD type I transmembrane glycoprotein. Nectin, originally isolated as poliovirus receptor-related protein (PRR), is a cell-cell adhesion molecule of the immunoglobulin supergene family. Nectins are calcium- independent immunoglobulin-like cell-cell adhesion molecules consisting of four members, nectin 1-4. Nectins homophilically and heterophilically trans-interact to form a variety of cell-cell junctions, including cadherin-based adherens junctions in epithelial cells and fibroblasts in culture, synaptic junctions in neurons, and Sertoli cell-spermatid junctions in testis, in cooperation with, or independently of, cadherins. Both nectin-2 and nectin-3 are ubiquitously expressed, whereas nectin-1 is abundantly expressed in brain. Nectin-2 and -3 are expressed in cells where cadherin is not expressed, such as blood cells and spermatids. All members of the nectin family have two or three splice variants. For nectin-3, three isoforms exist: nectin-3a, -3b and -3g of which nectin-3a is the largest. Nectin-3, also known as PRR3, is a transmembrane protein that is predominantly expressed in testis and placental tissues as well in many cell lines. Nectin interacts in vivo with both long and short isoforms of afadin, an actin binding protein, at cadherin-based cell-cell adherence junctions in various tissues and cell lines. Furthermore, the ectodomains of nectin-3 and CD155 (Poliovirus Receptor) have shown strong affinity to each other. Injection of this antibody into lumen of seminiferous tubules leads to disruption of the actin filaments in Sertoli cells at the Sertoli-maturing spermatid ectoplasmic specialization and exfoliation of maturing spermatids form the seminiferous epithelium.
Applications:
Suitable for use in Flow Cytometry, Functional Studies, ELISA, Immunoprecipitation, Immunofluorescence and Immunohistochemistry. Other applications not tested.
Recommended Dilution:
Immunohistochemistry: Frozen (PFA or acetone/methanol fixation)
Flow Cytometry: 1:50
Immunofluorescence: fixed in 2% paraformaldehyde in PBS optionally followed by methanol/aceton fixation
Functional Studies: injection of 1ug into lumen of seminiferous tubules
ELISA: Indirect ELISA on 0.5ug/ml mouse nectin-3 coated per well. Positive signal with 2ug/ml antibody.
Immunoprecipitation: 1-2ug per 100-500ug of total protein (1ml of cell lysate)
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20 degrees C. Aliquots are stable for 12 months at -20 degrees 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. FITC conjugates are sensitive to light.