Abstract:OsCR4 is the ortholog of ZmCR4 in rice and belongs to the family of TNFR (tumor necrosis factor receptor) receptor-like kinase in rice. Although ZmCR4 in maize, ACR4 in Arabidopsis have been reported to play important roles in growth and development, OsCR4 has been remained unidentified. Functional investigation of OsCR4 need anti-OsCR4 antibody. Here we depicted the preparation of anti-OsCR4 antibody. First, the antigen index of OsCR4 was analyzed with the program of DNA Star 5.0. A specific fragment in extracellular region of OsCR4 was chosed as an antigen section and cloned as fusion to the GST. Second, the recombinant proteins were produced by a large scale induction, GST affinity purification, SDS-PAGE separation and gel excision. Third, we immunized New Zealand rabbit with purified fusion proteins and acquired the polyclonal antibody with high titer of 1:512,000. What's more, this antibody could specially recognize the OsCR4 protein in microsomal fraction of rice leaves.
马云,孙大业,孙颖. 水稻类受体激酶OsCR4的抗体制备及特异性检测[J]. 中国生物化学与分子生物学报, 2006, 22(11): 924-930.
MAYun,SUNDa-ye,SUNYing. Preparation and identification of antibody to OsCR4, the receptor-like kinase from rice. Chinese Journal of Biochemistry and Molecular Biol, 2006, 22(11): 924-930.