Identification of CRISP2 from human sperm as PSP94‐binding protein and generation of CRISP2‐specific anti‐peptide antibodies

In this study, we identified CRISP2 as a potential binding protein of PSP94 from human sperm. Further, we generated antisera capable of specifically detecting CRISP2 and not CRISP3. In this direction, specific peptides corresponding to the least conserved ion channel regulatory region were synthesized, and polyclonal antibodies were generated against the peptide in rabbits. The binding characteristics of the anti‐CRISP2 peptide antibody were evaluated using competitive ELISA. Immunoblotting experiments also confirmed that the peptide was able to generate antibodies capable of detecting the mature CRISP2 protein present in human sperm lysate. Furthermore, this anti‐CRISP2 peptide antibody also detected the presence of native CRISP2 on sperm.Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

Synthesis of CRISP2‐specific peptides corresponding to the least conserved ion channel regulatory region, followed by production and characterization of anti‐CRISP2 peptide antibodies capable of specifically differentiating CRISP2 from CRISP3 and detecting native CRISP3 present on the sperm

Source: Journal of Peptide Science - April 30, 2016 Category: Biochemistry Authors: Jenifer H. Anklesaria, Bhalchandra J. Kulkarni, Bhakti R. Pathak, Smita D. Mahale Tags: Research Article Source Type: research