Nicotinamide N-methytransferase (NNMT) protein monoclonal antibody, hybridoma cell strain and application
A hybridoma cell line, monoclonal antibody technology, applied in the direction of anti-enzyme immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problems affecting the effect of radiation and radiotherapy, and achieve the effect of high antibody titer
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[0030] Example 1: Acquisition of Hybridoma Cell Lines, Screening and Preparation of Monoclonal Antibodies
[0031] 1. Preparation of monoclonal antibodies
[0032] 1.1. Preparation of immunogens
[0033] The immunogen is GST-NNMT fusion protein and NNMT protein. NNMT protein was prepared by genetic engineering technology: pGEX-4T-2 / NNMT plasmid was constructed and transformed into Escherichia coli BL-21-STAR (DE3) to express GST-NNMT fusion protein . Then, after the fusion protein was purified, the fusion protein was fully digested by adding thrombin, and the NNMT protein (geneBank accession number: ACA06031.1) was obtained after purification again.
[0034] 1.2. Animal immunity:
[0035] The immunization procedure was as follows: 6-8 week-old BALB / C female mice were taken for the initial immunization, NNMT protein and Freund's complete adjuvant were mixed in equal volumes, and then injected subcutaneously on the back of the mice at multiple points, with a dose of 120 μg pe...
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[0082] Example 2: Application of monoclonal antibodies to immunohistochemical detection of clinical specimens (two-step indirect method)
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