The monoclonal antibody 103-A1 recognizes mouse nectin-3. Nectin-3 is a 83 kDa 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 calciumindependent
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-3α, -3β and -3g of which nectin-3α 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 antibody 103-A1 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.
Flow cytometry, Frozen sections, Functional studies, Immuno assays, Immuno fluorescence
IF: fixed in 2% paraformaldehyde in PBS optionally followed by methanol/aceton fixation (Ref 2-7).
FC: 1 μg per 1 x 106 cells.
FS: injection of 1 μg into lumen of seminiferous tubules.
IA: Indirect ELISA on 0.5 μg/ml mouse nectin-3 coated per well. Positive signal with 2 μg/ml antibody.
IF: Cells are fixed in 2 % paraformaldehyde in PBS, optionally followed by methanol/aceton
fixation (Ref 1-6).
Dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
L-cells (A cultured line of C3H mouse fibroblasts)
Cells/tissues derived from nectin-3 knockout mouse (Ref 5).