Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Murine Anti-cardiolipin Antibodies
酶联免疫吸附法(ELISA)定量检测鼠抗心磷脂抗体
发布: 2012年06月20日第2卷第12期 DOI: 10.21769/BioProtoc.227 浏览次数: 13511
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Abstract
The circulating anticardiolipin antibody is a hallmark of antiphospholipid syndrome. It also appears in a number of autoimmune mouse models and is indicative of the break of tolerance against self antigens. This protocol describes a reliable method to determine the relative serum titer of anticardiolipin in autoimmune mouse models.
Keywords: Mouse (小鼠)
ELISA (ELISA)
Autoimmune (自身免疫性)
Anti-cardiolipin antibody (抗心磷脂抗体)
Materials and Reagents
- Cardiolipin (Sigma-Aldrich, catalog number: C0563 )
- Ethanol
- Phosphate buffered saline (PBS)
- Tween 20
- Na2HPO4 (anhydrous)
- NaH2PO4 (anhydrous)
- NaCl
- Fetal bovine serum (FBS) (Hyclone)
- Bovine serum albumin (BSA)
- Horseradish peroxidase (HRP) conjugated goat anti-mouse isotype specific antibodies [Southern Biotech, catalog number: 1040-05 (IgA); 1030-05 (IgG); 1021-05 (IgM)]
- ABTS Peroxidase Substrate Solution A and B (Kirkegaard & Perry Laboratories, catalog number: 50-62-01 )
- ABTS Peroxidase Stop Solution (Kirkegaard & Perry Laboratories, catalog number: 50-85-01 )
- 10x PBS-Tween 20 (see Recipes)
- Blocking solution (see Recipes)
Equipment
- Standard bench-top centrifuge
- Immulon 2HB plates (Fisher Scientific, catalog number: 14-245-61 )
- ELISA reader
Procedure
文章信息
版权信息
© 2012 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Liu, Z. (2012). Quantitative Enzyme-linked Immunosorbent Assay (ELISA) to Measure Serum Levels of Murine Anti-cardiolipin Antibodies. Bio-protocol 2(12): e227. DOI: 10.21769/BioProtoc.227.
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分类
免疫学 > 抗体分析 > 抗体检测
生物化学 > 蛋白质 > 免疫检测
免疫学 > 动物模型 > 小鼠
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