Human/Mouse/Rat Contactin‑1 Antibody

Cell Adhesion Mediated by Contactin‑1 and Neutralization by Human Contactin‑1 Antibody.

Recombinant Human Contactin-1 Fc Chimera (Catalog # 904-CN), immobilized onto a microplate, supports the adhesion of the C6 rat glioma cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human Contactin-1 Fc Chimera (2 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human/Mouse/Rat Contactin-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF904). The ND₅₀ is typically 1-5 µg/mL.

Detection of Contactin-1 by Immunohistochemistry

Expression and localization of Cntn1 in the retina.A–C In situ hybridization for Cntn1 expression in the wild type mouse retina at P5 (A), P10 (B), and P21 (C). A subset of cells in the retinal ganglion cell layer (bottom) and inner nuclear layer are positive for Cntn1 expression. Photoreceptors in the outer nuclear layer (top) do not have signals above background. D) Double label in situ hybridization with Cntn1 and syntaxin1a at P21 demonstrates that some amacrine cells in the inner nuclear layer are positive for Cntn1 expression (arrowheads). Other Cntn1-positive cells are likely to be bipolar cells based on their position (arrows). E) In the retinal ganglion cell layer, a majority of cells expressing Cntn1 at P21 also express Thy1, a marker of ganglion cells. F) Immunolabeling of retinas with anti-CNTN1 antibodies at P14 revealed strong labeling of the synaptic plexiform layers, as well as immunoreactivity in the cellular layers, particularly the inner nuclear layer. G) Immunolabeling retinas from Cntn1 mutant mice revealed a marked reduction, but not an elimination of signal intensity in images collected with equivalent parameters. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/22242131), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Contactin-1 by Immunohistochemistry

Expression and localization of Cntn1 in the retina.A–C In situ hybridization for Cntn1 expression in the wild type mouse retina at P5 (A), P10 (B), and P21 (C). A subset of cells in the retinal ganglion cell layer (bottom) and inner nuclear layer are positive for Cntn1 expression. Photoreceptors in the outer nuclear layer (top) do not have signals above background. D) Double label in situ hybridization with Cntn1 and syntaxin1a at P21 demonstrates that some amacrine cells in the inner nuclear layer are positive for Cntn1 expression (arrowheads). Other Cntn1-positive cells are likely to be bipolar cells based on their position (arrows). E) In the retinal ganglion cell layer, a majority of cells expressing Cntn1 at P21 also express Thy1, a marker of ganglion cells. F) Immunolabeling of retinas with anti-CNTN1 antibodies at P14 revealed strong labeling of the synaptic plexiform layers, as well as immunoreactivity in the cellular layers, particularly the inner nuclear layer. G) Immunolabeling retinas from Cntn1 mutant mice revealed a marked reduction, but not an elimination of signal intensity in images collected with equivalent parameters. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/22242131), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Contactin-1 by Western Blot

CNTN1 facilitates USP25 to degrade MAVS through the proteasomal pathway.(A) USP25 inhibits K63-linked ubiquitination of MAVS. HEK293 cells transfected with Myc-MAVS, HA-ubiquitin, or its mutants (K48O, K63O), together with a control or USP25 plasmids, were pre-treated with MG132 (10 μM) for 6 h, and the cells were then subjected to denature-IP and immunoblotting analysis with the indicated antibodies. (B) USP25+/+ and USP25-/- cells transfected with Myc-MAVS, HA-ubiquitin (K63O), were left uninfected or infected with SeV for 12 h. The cells were then subjected to denature-IP and immunoblotting analysis with the indicated antibodies. (C) USP25+/+ and USP25-/- cells were left uninfected or infected with SeV for 12 h. Twenty-four hours later, cells were harvested for SDD-AGE or SDS-PAGE followed by immunoblotting analysis with the indicated antibodies. (D) USP25-knockdown inhibits CNTN1-triggered K63-linked ubiquitination of MAVS. HEK293 cells were transfected with the indicated plasmids and USP25 siRNA or NC for 24 h. The cells were then treated with MG132 for 6 h before being used in ubiquitination assays with the indicated antibodies. (E) CNTN1+/+ or CNTN1-/- HEK293 cells were transfected with Myc-MAVS and HA-ubiquitin (K63O), together with a control or USP25 plasmids. The cells were then subjected to denature-IP and immunoblotting analysis with the indicated antibodies. (F) USP25-knockdown inhibits CNTN1-mediated degradation of MAVS. HEK293 cells were transfected with the indicated plasmids and USP25 siRNA or NC for 24 h before immunoblotting analysis. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35171955), licensed under a CC-BY license. Not internally tested by R&D Systems.