A kind of monoclonal antibody against human btf and its application
A monoclonal antibody, species-specific technology, applied in the field of antibody engineering and medicine, can solve problems such as poor stability and chromosome deletion
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Embodiment 1
[0016] Example 1 Preparation and Screening of Mouse Monoclonal Antibody
[0017] (1) Antigen preparation
[0018] The human BTF (Bcl-2-associated transcription factor 1) gene was amplified from the human gene transcriptome by PCR. The upstream primer was: 5'-GGCCGCTAGCATGGACTACAAGGACGACGATGACAAGGGTCGCTCCAATTCTAG-3'; the downstream primer was: 5'-GGCCGGATCCTTATTCCTTTTCTTCCTTGC-3'. Using NheⅠ and BamHI double enzyme digestion, the gene fragment was connected into the eukaryotic expression vector pIRESneo3, PCR and double enzyme digestion identified positive clones and then sequenced. After the sequence was determined correctly, the plasmid was extracted, and Lipofectamine2000 (Invitrogen) mediated transfection into 293T cells, G418 Screen stable clones and transfer to culture flasks for expression. Cells were collected by centrifugation, non-denatured cells were lysed, centrifuged at 10,000rpm for 10 minutes, BTF was extracted by FLAG affinity chromatography, the eluate was use...
Embodiment 2W
[0028] Embodiment 2Western-Blot detects the expression of BTF
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