Nonradioactive labeling immunoprecipitation method for detecting NXP2 autoantibody of inflammatory myopathies and application
An autoantibody and immunoprecipitation technology, which is applied in the direction of viruses/bacteriophages, tumor-specific antigens, chemical instruments and methods, etc., can solve problems such as high false positive rate and high cost of ELISA kits
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0023] Example 1 Non-radiolabeled immunoprecipitation detection method of NXP2 autoantibodies in inflammatory myopathy
[0024] A non-radiolabeled immunoprecipitation detection method for NXP2 autoantibodies in inflammatory myopathy, comprising the following steps:
[0025] (1) Design primer pair NXP2-F (5'-CGGTCGACCGCGGCGCAGCACC-3') and NXP2-R2 (5'-GCGGTACCTTAGCTTGGAGTTGAAAGAGC-3'), primer pair NXP2-F2 (5'-CGGTCGACCGAAGATACAGAAGCGTCC-3') and NXP2-R (5'-GCGGTACCTTAAGTACTACTGATTTCACTCATT-3'), and then the N-terminal (Genbank: NM_015358.2, 80-1576bp) or C-terminal (Genbank: NM_015358.2, 1274-2896bp).
[0026] (2) Recover and purify the PCR fragment with a gel recovery kit (Omega, D2500-01), perform double digestion with SalI and KpnI restriction endonucleases, and ligate overnight with the pCMV-myc vector that has been double-digested with SalI and KpnI , Transform JM109 bacteria competent, screen positive clones on a plate containing 50ug / ml ampicillin, extract a small amount...
Embodiment 2
[0030] Example 2 detects the positive rate of NXP2 antibody in serum of 50 patients with inflammatory myopathy
[0031] (1) Inoculate 30% HEK293 cells on 75cm 2 Cell culture dish, cultivated at 37°C for less than 24 hours;
[0032] (2) Transfect HEK293 cells with 20ug pCMV-myc-NXP2C plasmid for 48 hours;
[0033] (3) HEK293 cells were lysed with 1 ml of RIPA lysate containing protease inhibitors, and the protein was quantified;
[0034] (4) Using c-myc antibody to confirm the overexpressed NXP2 C-terminus in HEK293 cells.
[0035] Take 20ul of the test serum, positive serum, and negative serum from patients with inflammatory myopathy for immunoprecipitation with the cell lysate overexpressing the C-terminus of NXP2, perform SDS-PAGE gel electrophoresis, and then detect with c-myc antibody to determine the presence of NXP2 antibody in The positive rate in patients with inflammatory myopathy was about 12.8% (as attached Figure 6 As shown, (a) and (b) are respectively select...
PUM

Abstract
Description
Claims
Application Information

- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com