Reagent kit for detecting autoimmunity disease related antinuclear antibodies spectrum and preparation method thereof
An autoimmune disease and anti-nuclear antibody spectrum technology, applied in the field of biomedicine, can solve problems such as misjudgment of results, destruction of antigenic determinant conformation, non-specific binding of non-specific antibodies, etc., to achieve simple operation, improved sensitivity, The effect of less sample consumption
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Example Embodiment
[0034] Example 1 Preparation of the kit of the present invention
[0035] 1. Preparation of natural antigen
[0036](1) Natural antigen extraction: nRNP / Sm, Sm, SS-A, SS-B, Scl-70, Jo-1, dsDNA, nucleosomes, histones and ribosomal P protein are extracted from calf thymus by acetone method Natural protein extracted from tissue; AMA-M2 natural protein extracted from porcine heart tissue. The specific method is as follows:
[0037] Calf thymus and pig's heart are stored in an ultra-low temperature refrigerator at -80°C. First, ANA is extracted with phosphate buffered saline (PBS) and Tris-HCl buffer at 4°C, and then precipitated with acetone pre-cooled at -20°C. Extract ANA into dry powder, do polyacrylamide gel electrophoresis (SDS-PAGE) and vertical transfer (Western blot) together with Sigma rabbit thymus acetone powder, transfer to nitrocellulose membrane, ID and IBT detect anti-ANA antibody, Secondly, different saturated concentrations of ammonium sulfate precipitated pig heart a...
Example Embodiment
[0049] Example 2 Method of using the kit of the present invention
[0050] Sample requirements: 2-5ml of intravenous blood, placed in a test tube to be coagulated and centrifuged to separate the serum. Serum samples are stored at 2-8°C. Specimens that cannot be tested within 24 hours should be placed at -20°C, severely hemolyzed, flocculent or mildew The changed serum sample will affect the test.
[0051] 1. Add samples
[0052] Dilute the sample to be tested with the sample diluent at a ratio of 1:100, add 100ul to the reaction tank of the detection reagent prepared in the above-mentioned embodiment 1, and put it into the incubation shaker. React at 37°C for 30 minutes.
[0053] 2. Wash the board
[0054] After the sample reaction is over, discard the reaction solution, shake and wash for 3×5 min.
[0055] 3. Add enzyme-labeled secondary antibody
[0056] Alkaline phosphatase-labeled goat anti-human IgG antibody was diluted 10 times, 100ul was added to each reaction tank, placed ...
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.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap