Production of polyclonal antibody against human Neuritin and its application of immunodetection

Abstract

Objective: To date, a commercial antibody to human Neuritin for immunoprecipitation is still limited. In this study, we aimed to develop a specific antibody for further research on the potential function of Neuritin.

Methods and results: By epitope prediction of recombinant human Neuritin, the active fragment of human Neuritin that could be used as an excellent immunogen. Soluble His-tagged Neuritin was expressed and purified from Pichia pastoris. Polyclonal antibody against Neuritin was obtained by immunizing Sprague-Dawley rats with purified recombinant human Neuritin. Affinity-purified polyclonal antibody against Neuritin was characterized with indirect enzyme-linked immunosorbent assay, immunoblotting, immunoprecipitation, and immunofluorescence. The results demonstrated that the polyclonal antibody against Neuritin had been prepared successfully. The prepared antibody bound to both exogenous and endogenous Neuritin. Importantly, the anti-Neuritin polyclonal antibody could be used in immunoprecipitation assays.

Conclusions: The prepared polyclonal antibody could be used in immunoprecipitation and provide researchers with a useful tool for further investigating the function and mechanism of Neuritin.

Keywords:

• Antiserum
• immunofluorescence
• immunoprecipitation
• neuritin
• ployclonal antibody

Acknowledgments

We thank Dr. Tiepeng Wang for providing SH-SY5Y cells and Pan Zhang for offering help.

Additional information

Funding

This work was supported by the National Natural Science Foundation under Grant number 81771173; Shihezi University High-Talents Funding under Grant number RCZX200676.