Preparation and identification of polyclonal antibody against LRR-1 protein of Chlamydomonas reinhardtii
Chlamydomonas reinhardtii was used as a model organism to deeply study the function mechanism of leucine rich repeat(LRR)in the process of innate immunity in the organism.The physico-chemical properties of CrLRR-1 protein including secondary structure,hydrophilicity,and immunogenicity were predicted and analyzed with bioinformatics.The CrLRR-1 specific peptide was artificially designed and synthesized,then coupled with keyhole limpet hemocyanin(KLH)to immunize New Zealand white rab-bits.The results showed that the antiserum titer was determined to be 1∶512 000 by indirect ELISA.The CrLRR-1 protein polyclonal antibodies were obtained after sequentially purifying the antiserum with Protein A and antigen affinity.The antibodies prepared with immunoblotting and immunofluorescence detection spe-cifically recognized the endogenous LRR-1 protein in Chlamydomonas reinhardtii,and the CrLRR-1 pro-tein was mainly localized on the cell membrane.It is indicated that the polyclonal antibody against CrLRR-1 protein were successfully prepared and identified.
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