Production, selection and characterization of monoclonal antibodies against AHP1 protein that has function in Arabidopsis Thaliana signaling pathway

In the present study; production, selection and characterization of monoclonal antibodies against AHP1 protein that has function in Arabidopsis thaliana signaling pathway are completed. Monoclonal antibodies are purified by using Ni Sepharose high performance chromatography column. Monoclonal antibodies were characterized by using ELISA and Western Blot techniques and three of them are classified as monospecific and two of them classified as polyspecific. Using by isotyping method; monoclonal antibodies are tested on IgG1, IgG2a, IgG2b, IgG3, IgGM and IgGA (secondary antibodies) to find out which secondary antibody gives best reaction with antibodies. Antibodies are tested on crude extracts and as a result; two of the monospecific antibodies are only specific to AHP1 protein, other monospecific antibody is specific to AHP1 and has cross reaction with AHP6 and polyspecific antibodies are specific to AHP1,2,3,4,5,6 proteins. Sensitivity test is used for each antibodies and best dilutions are calculated. Besides these epitope mapping is completed. According to MS/MS analysis; AHP1 protein is bounded to one of the antibody on KVDPHVHQLK epitope.