Technical datasheet
Description
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 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-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.
Cross Reactivity
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Cross reactant
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Reactivity
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Human nectin-3
Rat nectin-3
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No
No
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Immunogen
Recombinant mouse nectin-3 extracellular domain (50-400 aa) fused to IgG Fc
Formulation
1 ml (100 µg/ml) 0.2 µm filtered FITC conjugated antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide
Species
Rat IgG2a
Application
| | F2-7 | FC | FS7 | IA | IF1-6 | IP | P | W | | Yes | ● | ● | ● | ● | ● | ● | | | | No | | | | | | | | | | N.D. | | | | | | | ● | ● | |
N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot Application IA has been tested by Hycult Biotech. Application FC, IP are based on personal communication. |
Application notes
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 (Ref 7)
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)
IP: 1-2 μg per 100-500 μg of total protein (1 ml of cell lysate)
Use
For immunohistochemistry and flow cytometry, 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.
Aliases
PPR3, poliovirus receptor-related protein 3, PRR3, PVRR3, CD113
Positive control
L-cells (A cultured line of C3H mouse fibroblasts),
Negative control
Cells/tissues derived from nectin-3 knockout mouse (Ref 5)
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
References
1. Satoh-Horikawa, K et al; Nectin-3 a new member of immunoglobulin-like cell adhesion molecules that shows homophilic and heterophilic cell-cell adhesion activities. J Biol Chem 2000, 275: 10291
2. Mizoguchi, A et al; Nectin: an adhesion molecule involved in formation of synapses. J Cell Biol, 2002, 156: 555
3. Mueller, S et al; Loss of nectin-2 at Sertoli-spermatid junctions leads to male infertility and correlates with severe spermatozoan head and midpiece malformation, impaired binding to the zona pellucida, and oocyte penetration. Biol Reprod 2003, 69: 1330
4. Guttman, J et al; Evidence that tubulobulbar complexes in the seminiferous epithelium are involved with internalization of adhesion junctions. Biol of Reprod 2004, 71: 548
5. Inagaki, M et al; Roles of cell-adhesion molecules nectin 1 and nectin 3 in ciliary body development. Development 2005, 132: 1525
6. Inagaki, M et al; Role of cell adhesion molecule nectin-3 in spermatid development. Genes Cells,2006, 11: 1125
7. Toyama, Y et al; Disruption of ectoplasmic specilizations between Sertoli cells and maturing spermatids by anti-nectin-2 and anti-nectin-3 antibodies. Asain J Androl 2008, 10: 577
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
Also available
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HM1052
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Nectin-2, Mouse, mAb 502-57
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HM3013
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L-Afadin, Rat, mAb 3
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HM1050
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JAM-A, Mouse, mAb BV12
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HM1056
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JAM-C, Mouse, mAb 19H36
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HM1057
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JAM-C, Mouse, mAb CRAM-18F26
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References
1. Satoh-Horikawa, K et al; Nectin-3 a new member of immunoglobulin-like cell adhesion molecules that shows homophilic and heterophilic cell-cell adhesion activities. J Biol Chem 2000, 275: 10291
2. Mizoguchi, A et al; Nectin: an adhesion molecule involved in formation of synapses. J Cell Biol, 2002, 156: 555
3. Mueller, S et al; Loss of nectin-2 at Sertoli-spermatid junctions leads to male infertility and correlates with severe spermatozoan head and midpiece malformation, impaired binding to the zona pellucida, and oocyte penetration. Biol Reprod 2003, 69: 1330
4. Guttman, J et al; Evidence that tubulobulbar complexes in the seminiferous epithelium are involved with internalization of adhesion junctions. Biol of Reprod 2004, 71: 548
5. Inagaki, M et al; Roles of cell-adhesion molecules nectin 1 and nectin 3 in ciliary body development. Development 2005, 132: 1525
6. Inagaki, M et al; Role of cell adhesion molecule nectin-3 in spermatid development. Genes Cells,2006, 11: 1125
7. Toyama, Y et al; Disruption of ectoplasmic specilizations between Sertoli cells and maturing spermatids by anti-nectin-2 and anti-nectin-3 antibodies. Asain J Androl 2008, 10: 577