Native BAD-1 Binding to Heparin-agarose
天然BAD-1与肝素琼脂糖的结合
发布: 2014年04月05日第4卷第7期 DOI: 10.21769/BioProtoc.1095 浏览次数: 7184
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参见作者原研究论文
The authors used this protocol in:
Jul 2013
Abstract
BAD-1 is an adhesin created by the dimorphic fungus Blastomyces dermatitidis, the causative agent of blastomycosis. We have determined that it has an affinity for heparin, which may explain its impact on virulence and human immune function as a number of cells related to immune function have heparin like moieties on their surfaces. This assay allows a quantification of binding between soluble BAD-1 and immobilized heparin.
Materials and Reagents
- Heparin-agarose resin (Sigma-Aldrich, catalog number: H6508 ) (prior to use it is washed 3x in five volumes of tricine buffer to eliminate free heparin)
- 10 μg of BAD-1 [purified according to the method of Brandhorst et al. (2005)]
- 25 microliters of soluble medical-grade sodium heparin for injection (50 mg/ml) (Elkins-Sinn Inc)
- Tricine buffer (Sigma-Aldrich, catalog number: T0377 ) (see Recipes)
Equipment
- Accuspin micro17 microcentrifuge (Thermo Fisher Scientific)
- Nanodrop ND1000 spectrophotometer
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Brandhorst, T. T. (2014). Native BAD-1 Binding to Heparin-agarose. Bio-protocol 4(7): e1095. DOI: 10.21769/BioProtoc.1095.
- Taguchi, Y., Mistica, A. M., Kitamoto, T. and Schätzl, H. M. (2013). Critical significance of the region between Helix 1 and 2 for efficient dominant-negative inhibition by conversion-incompetent prion protein. PLoS Pathog 9(6): e1003466.
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分类
微生物学 > 微生物生物化学 > 蛋白质
生物化学 > 蛋白质 > 相互作用
生物化学 > 蛋白质 > 活性
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