成年小鼠脑切片的X-gal染色

Hiroshi  Kokubu; Janghoo  Lim

doi:10.21769/BioProtoc.1064

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Peer-reviewed

X-gal Staining on Adult Mouse Brain Sections

成年小鼠脑切片的X-gal染色

HK Hiroshi Kokubu

Janghoo Lim email

发布: 2014年03月05日第4卷第5期 DOI: 10.21769/BioProtoc.1064 浏览次数: 17951

评审: Hui ZhuXuecai Ge

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参见作者原研究论文

The authors used this protocol in:

Cover of The Journal of Neuroscience, featuring study using the protocol.

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

English

中文翻译

文章信息

版权信息

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.
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.