Preparation And Application Of Rabbit Anti-osteoclastic Protein-tyrosine Phosphatase Polyclonal Antibody

In clinical orthodontic treatment,the alveolar bone is remodeled under the induction of orthodontic force,then the teeth are moved.However,on the pressure side,interleukins,tumor necrosis factor alpha and other factors are produced in the periodontal tissues.Under the action of a series of signal transduction factors,different degrees of absorption occur on the root surface of the pressure side.At present,the mechanism of root resorption and development in the orthodontic community is not yet fully understood and how to avoid and reduce the degree of root resorption is an issue that every orthodontist should pay attention to it during the orthodontic treatment.Protein tyrosine phosphatases(Protein Tyrosine Phosphatases,PTPs)are a kind of intracellular regulatory factors involved in many cell signal transduction processes.Their regulatory effects on osteoclasts have gradually been discovered by people in recent years.PTPs are involved in the differentiation,proliferation,activation and maturation of bone cells.In recent years,a large number of studies have shown that Osteoclastic protein-tyrosine phosphatase(PTP-oc)plays a significant regulatory role in the differentiation,formation,and regulates the activity of osteoclasts.At the same time,PTP-oc is an important regulator of bone metabolism in vivo.Therefore,an in-depth study of the function of PTP-oc will provide experimental evidence for the prevention and treatment of orthodontic root resorption in the future.For this reason,the rabbit anti-polyclonal antibody against PTP-oc was prepared for the first time by immunizing rabbits,which laid the foundation for further research on the function and mechanism of PTP-oc.The constructed p ET28a-ΔPTP-oc recombinant plasmid was transformed into E.coli BL21(DE3),and the Isopropyl-β-D-thiogalactoside(IPTG)was used to induce the expression of protein in 16℃,24 h.The recombinant protein was purified by the method of Q fast flow agarose and Ni-NTA agarose,and the purified recombinant protein was used as an antigen to immunize the rabbit to obtain anti-PTP-oc polyclonal antibody.The 4th immune blood was identified by the indirect ELISA.The indirect ELISA results showed that after immunizing rabbit with the recombinant PTP-oc protein,the polyclonal antibody with the titer of 1:32 000 was gotten.The Western blotting results showed that the polyclonal antibody possessed high specificity.