X-gal Staining on Adult Mouse Brain Sections
成年小鼠脑切片的X-gal染色
发布: 2014年03月05日第4卷第5期 DOI: 10.21769/BioProtoc.1064 浏览次数: 18802
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参见作者原研究论文
The authors used this protocol in:
May 2013
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Abstract
Knowing expression patterns of given proteins is very important to understand their functions. Immunostaining analysis with specific antibodies is commonly used to identify cells or tissues expressing proteins of interest. Although this technique is regularly used, it requires high quality of specific antibodies and there is no good quality of antibody available for certain proteins. Alternatively, X-gal staining is also used to analyze protein expression pattern. It is simple and routinely used to detect expression pattern of any proteins of interest in vivo. In this method, genetically modified animals that express beta-galactosidase under the control of certain regulatory elements will be used to reveal the expression pattern of proteins that use the same regulatory elements.
Keywords: Adult mouse brain (成年小鼠脑)
X-gal staining (X-gal染色)
Expression (表达)
Materials and Reagents
- Adult mouse brain
- Paraformaldehyde (PFA) (Sigma-Aldrich, catalog number: P6148 )
- Phosphate buffered saline (PBS)
- Sucrose
- Magnesium chloride, 6-Hydrate (MgCl2.6H2O) (Mallinckrodt Baker, catalog number: 2444-01 )
- Sodium deoxycholate (Sigma-Aldrich, catalog number: D6750 )
- NP-40 (Sigma-Aldrich, catalog number: I3021 )
- Potassium Ferricyanide (ACROS ORGANICS, catalog number: 196785000 )
- Potassium Ferrocyanide (Mallinckrodt Baker, catalog number: 6932-04 )
- Permount (Fisher Scientific, catalog number: SP15-100 )
- O.C.T. compund (Sakura, catalog number: 4583 )
- Ethanol (Decon Labs, catalog number: 2716 )
- X-gal (American Bioanatycal, catalog number: AB00450-00005 )
- 1 M MgCl2 (see Recipes)
- 10% Sodium deoxycholate (see Recipes)
- 20% NP-40 (see Recipes)
- 50 mM Potassium Ferricyanide (see Recipes)
- 50 mM Potassium Ferrocyanide (see Recipes)
- Staining buffer (see Recipes)
Equipment
- Cryomold (Sakura, catalog number: 4557 )
- Cryostat
- 37 °C incubator
- 10-20 Slide Staining Dish with Cover (VWR International, catalog number: 900203 ) or equivalent
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:
- Kokubu, H. and Lim, J. (2014). X-gal Staining on Adult Mouse Brain Sections. Bio-protocol 4(5): e1064. DOI: 10.21769/BioProtoc.1064.
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Ju, H., Kokubu, H., Todd, T. W., Kahle, J. J., Kim, S., Richman, R., Chirala, K., Orr, H. T., Zoghbi, H. Y. and Lim, J. (2013). Polyglutamine disease toxicity is regulated by Nemo-like kinase in spinocerebellar ataxia type 1. J Neurosci 33(22): 9328-9336.
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分类
神经科学 > 神经解剖学和神经环路 > 动物模型
生物化学 > 蛋白质 > 表达
细胞生物学 > 组织分析 > 组织分离
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