Monoclonal antibody of anti-human BAP31 protein, preparation method and application thereof
A monoclonal antibody and protein technology, applied in anti-animal/human immunoglobulin, botanical equipment and methods, biochemical equipment and methods, etc. Check the effect of the tool
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[0027] 1. Preparation of MCF-7 cell monoclonal antibody library
[0028] (1) Material method
[0029] 1. Source of experimental materials
[0030] BALB / c mice, MCF-7 cells and mouse myeloma cells (SP2 / 0) were provided by Tianjin Cancer Hospital. Freund's complete adjuvant and incomplete Freund's adjuvant, HAT, and HT conditioned medium components were purchased from Invitrigen. DMEM medium, polyethylene glycol (PEG), and RPMI 1640 medium were purchased from Hyclone Company. Horseradish peroxidase (HRP)-labeled goat anti-mouse Ig, o-phenylenediamine (OPD), diaminobenzidine (DAB), and 8-azaguanine (8-AG) were purchased from Sigma.
[0031] 2. Animal immunization
[0032] Take 1×10 7 MCF-7 cells were collected by centrifugation, added 200ul normal saline and repeatedly frozen and thawed 5 times, and the supernatant was discarded by centrifugation. Add 300 ul of lysate (0.05M Tris-HCl pH=8.0, 1% Triton X100) to the precipitate, centrifuge at 12,000 rpm at 4°C, and take the ...
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