Human Anti-RTN4R Recombinant Antibody (M5) (CAT#: HPAB-0738-YJ)

Figure 1 Analysis of the specificity and species cross-reactivity of M5 and P14 Fab by ELISA.

Binding of M5 and P14 Fab to immobilized human (h) and mouse (m) NgR1, human and rat (r) NgR2, human and rat NgR3, and BSA was analyzed by ELISA using goat antihuman Fab polyclonal antibodies conjugated to horseradish peroxidase.

Hofer, T., Tangkeangsirisin, W., Kennedy, M. G., Mage, R. G., Raiker, S. J., Venkatesh, K., ... & Rader, C. (2007). Chimeric rabbit/human Fab and IgG specific for members of the Nogo-66 receptor family selected for species cross-reactivity with an improved phage display vector. Journal of immunological methods, 318(1-2), 75-87.

Figure 2 Analysis of the specificity and species cross-reactivity of M5 and P14 Fab by Western blotting.

For Western blotting, lysates of 293F cells transfected with vectors for mammalian cell expression of human NgR1, NgR2, and NgR3, respectively, were separated by SDS-PAGE, blotted onto nitrocellulose, and probed with M5 and P14 Fab followed by goat anti-human Fab polyclonal antibodies conjugated to horseradish peroxidase. Goat anti-human NgR3 polyclonal antibodies followed by donkey anti-goat polyclonal antibodies conjugated to horseradish peroxidase were used as positive control for the expression of human NgR3.

Hofer, T., Tangkeangsirisin, W., Kennedy, M. G., Mage, R. G., Raiker, S. J., Venkatesh, K., ... & Rader, C. (2007). Chimeric rabbit/human Fab and IgG specific for members of the Nogo-66 receptor family selected for species cross-reactivity with an improved phage display vector. Journal of immunological methods, 318(1-2), 75-87.

Figure 3 Flow cytometry analysis of M5 and P14 Fab binding to transfected human 293F cells.

Flow cytometry histograms showing the binding of M5 Fab (left) and P14 Fab (right) to 293F cells transfected with vectors for mammalian cell expression of human (h) NgR1, NgR2, and NgR3, respectively. FITC-conjugated goat anti-human Fab polyclonal antibodies were used for detection.

Hofer, T., Tangkeangsirisin, W., Kennedy, M. G., Mage, R. G., Raiker, S. J., Venkatesh, K., ... & Rader, C. (2007). Chimeric rabbit/human Fab and IgG specific for members of the Nogo-66 receptor family selected for species cross-reactivity with an improved phage display vector. Journal of immunological methods, 318(1-2), 75-87.

Figure 4 Immunofluorescence microscopy analysis of M5 and P14 IgG1κ binding to stably transfected rat PC12 cells.

Binding of M5 and P14 IgG1κ to rat PC12 cells stably transfected with vectors for mammalian cell expression of rat (r) NgR1 or NgR2 was detected with Alexa Fluor® 594-conjugated goat anti-human IgG polyclonal antibodies.

Hofer, T., Tangkeangsirisin, W., Kennedy, M. G., Mage, R. G., Raiker, S. J., Venkatesh, K., ... & Rader, C. (2007). Chimeric rabbit/human Fab and IgG specific for members of the Nogo-66 receptor family selected for species cross-reactivity with an improved phage display vector. Journal of immunological methods, 318(1-2), 75-87.

Figure 5 Immunoprecipitation of exogenously and endogenously expressed rat and mouse NgR1.

Lysates from (A) rat PC12 cells stably transfected with vectors for mammalian cell expression of rat (r) NgR1 or NgR2 and (B) lysates from adult mouse cortex/hippocampus membranes were subjected to immunoprecipitation using M5 IgG1κ and Protein A/G beads. P14 IgG1κ were used as negative control. Input (IN), supernatants (SN), and immunoprecipitates (IP) were separated by SDS-PAGE, blotted onto nitrocellulose, and probed with goat anti-human NgR1 polyclonal antibodies conjugated to horseradish peroxidase.

Hofer, T., Tangkeangsirisin, W., Kennedy, M. G., Mage, R. G., Raiker, S. J., Venkatesh, K., ... & Rader, C. (2007). Chimeric rabbit/human Fab and IgG specific for members of the Nogo-66 receptor family selected for species cross-reactivity with an improved phage display vector. Journal of immunological methods, 318(1-2), 75-87.