A kind of serum cenpf antibody quantitative detection kit
A GST-CENPF, quantitative detection technology, applied in the field of serum CENPF antibody quantitative detection kits, can solve problems such as long-term clinical application
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Embodiment 1
[0019] Example 1 Preparation of CENPF recombinant protein
[0020] 1. Materials
[0021] DNA marker (100-2000bp) and rainbow prestained protein marker (14-120KD) were purchased from Beijing Tiangen Biochemical Technology Co., Ltd.; chemiluminescent protein marker (20-90KD) was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; Bacillus Origammi2 and BL21 (DE3) were preserved in our laboratory; Fast Taq Mastermix, BamHI, XhoI, and T4 DNA ligase were purchased from NEB Company; DNA electrophoresis gel recovery kit was purchased from Axygen Company; D-galactoside (IPTG) was purchased from Amresco; GST antibody was purchased from Beyond Biotechnology Research Institute; HRP-labeled goat anti-human IgG was purchased from Earthox, USA; primer synthesis and DNA sequence determination were provided by Shanghai Sangon Bioengineering Co., Ltd. Finish. All other reagents were of domestic analytical grade.
[0022] 2. Construction and identification of human CENPF prokaryotic e...
Embodiment 2
[0035] Example 2 Preparation of ELISA plate
[0036] 1. Materials
[0037] The 96-well ELISA plate was purchased from Thermo Company in the United States; the recombinant CENPF antigen protein was prepared in our laboratory; BSA was purchased from Beijing Huanya Tech Biomedical Technology Co., Ltd.; TMB color development solution and stop solution were purchased from Beijing Solaibao Technology Co., Ltd. company; HRP-labeled rabbit anti-human IgG was purchased from Sigma, USA; other reagents were of domestic analytical grade.
[0038] 2. Preparation of standard products in the detection kit
[0039] In the previous research process, our research group has quantitatively detected the serum CENPF antibody level of a group of liver cancer patients through protein chip technology, and screened 30 cases of serum containing high levels of CENPF antibody (protein chip detection signal value greater than 30,000) The serum samples were mixed as the standard of this test kit.
[0040...
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