成年小鼠脑切片的X-gal染色

Hiroshi  Kokubu; Janghoo  Lim

doi:10.21769/BioProtoc.1064

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
登录
/
注册
- 个人主页
- 编辑个人信息
- 修改密码
- 退出
CN
- EN - English
- CN - 中文

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: 18337

评审: Hui ZhuXuecai Ge

PDF

Q&A

引用

Cited by

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

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

Cell-Sonar：通过特定蛋白标志物表达变化追踪目标蛋白的简便低成本方法

Sabrina Brockmöller [...] Simone Rothmiller

2025年02月05日 1324 阅读

脑组织微血管的分离与成像

Josephine K. Buff [...] Sophia M. Shi

2025年08月05日 2141 阅读

Cluster FLISA——用于比较不同细胞系蛋白表达效率及蛋白亚基聚集状态的方法

Sabrina Brockmöller and Lara Maria Molitor

2025年11月05日 630 阅读

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.