LightShift™ EMSA 优化和控制试剂盒
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LightShift™ EMSA 优化和控制试剂盒

The Thermo Scientific LightShift EMSA Optimization and Control Kit is an extraordinarily robust and sensitive system for performing electrophoretic mobility了解更多信息
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货号数量
20148X100 次反应
货号 20148X
价格(CNY)
1,770.00
飞享价
Ends: 31-Dec-2025
2,113.00
共减 343.00 (16%)
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添加至购物车
数量:
100 次反应
请求批量或定制报价
价格(CNY)
1,770.00
飞享价
Ends: 31-Dec-2025
2,113.00
共减 343.00 (16%)
Each
添加至购物车
The Thermo Scientific LightShift EMSA Optimization and Control Kit is an extraordinarily robust and sensitive system for performing electrophoretic mobility shift assays (EMSA) to identify and characterize protein-DNA binding interactions. The kit includes reagents for setting up and customizing DNA binding reactions and a control set of DNA and protein extract to test the kit system.

Features of theLightShift EMSA Optimization and Control Kit:

• Excellent for detecting low-abundance proteins in nuclear extracts
• Sensitivity that surpasses radioactive and digoxigenin methods
• Compatible with previously established binding conditions for popular DNA-protein interactions
• Includes EBNA control system to help new users develop a working assay and understand the methods used to confirm binding interaction specificity

The principle for LightShift EMSA Detection is similar to a Western blot. Biotin end-labeled duplex DNA is incubated with a nuclear extract or purified factor and electrophoresed on a native gel. The DNA is then rapidly (30 minutes) transferred to a positive nylon membrane, UV crosslinked, probed with streptavidin-HRP conjugate and incubated with the substrate. The protocol from labeling to results can be accomplished in a single day.

The interaction of proteins with DNA is central to the control of many cellular processes including DNA replication, recombination and repair, transcription, and viral assembly. One technique that is central to studying gene regulation and determining protein:DNA interactions is the electrophoretic mobility shift assay (EMSA).

The EMSA technique is based on the observation that protein:DNA complexes migrate more slowly than free DNA molecules when subjected to non-denaturing polyacrylamide or agarose gel electrophoresis. Because the rate of DNA migration is shifted or retarded upon protein binding, the assay is also referred to as a gel shift or gel retardation assay. Until conception of the EMSA protein:DNA interactions were studied primarily by nitrocellulose filter-binding assays.

All that is needed to perform the assay is purified DNA target that has been end-labeled with biotin, the protein extract to be tested, nylon membrane and basic electrophoresis equipment. DNA targets can be synthesized with 5' or 3' biotin labels or they can be labeled after synthesis using the Thermo Scientific Biotin 3' End DNA Labeling Kit (Product No. 89818). Nuclear, cytosolic or whole cell protein extracts can be obtained by a variety of methods, including the Thermo Scientific NE-PER Nuclear and Cytoplasmic Extraction Reagent Kit (Product No. 78833).

More Product Data
Transfer EMSA gels using the Pierce G2 Fast Blotter

Related Products
LightShift™ Chemiluminescent EMSA Kit
LightShift™ Poly (dI-dC)
仅供科研使用。不可用于诊断程序。
规格
检测EMSA Assay
适用于(设备)myECL™ 成像仪、X 射线胶片
包括10X 结合缓冲液 (1 mL)、生物素-EBNA 对照 DNA (50 µL)、未标记 EBNA DNA (50 µL)、EBNA 提取物 (125 µL)、Poly dI·dC (125 µL)、50% 甘油 (500 µL)、1% NP-40 (500 µL)、1 M KCl (1 mL)、100 mM MgCl(500 µL)、200 mM EDTA(pH 值 8.0;500 µL)、5X 上样缓冲液 (1 mL)
产品线LightShift™
产品类型LightShift EMSA 优化和控制试剂盒
数量100 次反应
靶标特异性无靶标特异性
技术凝胶迁移
产品规格试剂盒
Unit SizeEach
内容与储存
足够用于:100 次结合反应
• 10X 结合缓冲液,1 mL
•生物素-EBNA 控制 DNA,50 µL
•未标记 EBNA DNA,50 µL
• EBNA 提取物,125 µL
• Poly (dI·dC),125 µL
• 50% 甘油,500 µL
• 1% NP-40,500 µL
• 1 M KCl,1 mL
• 100 mM MgCl2,500 µL
• 200 mM EDTA pH 值 8.0,500 µL
• 5X 上样缓冲液,1 mL

在 -20°C 下储存。

常见问题解答 (FAQ)

能否使用NativePAGE凝胶进行EMSA(电泳迁移率变动分析)?

NativePAGE凝胶已被成功用于EMSA,对2种纯化蛋白质间的相互作用进行分析。但是,我们尚未测试将NativePAGE凝胶用于EMSA以分析核酸(DNA或RNA)与蛋白质或蛋白质复合物间的相互作用。

If the LightShift Chemiluminescent EMSA kit has been improperly stored (i.e., at room temperature, -20°C or +4°C), will it still work correctly?

The LightShift Chemiluminescent EMSA Kit is composed of two sets of components that require different storage temperatures. One component set consists of the chemiluminescent substrates and various buffers that are stored at 4°C. The other component set consists of the control DNAs and various optimization reagents that are stored at -20°C. The EBNA extract must be maintained at -20°C or it will lose activity (proteins will degrade). Short-term storage (overnight) of the other kit components at temperatures ranging from room temperature to -20°C will not adversely affect kit performance.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I am using the LightShift Chemiluminescent EMSA kit. Can I probe for the proteins by performing a Western blot?

This has not been tested but may be possible. A better alternative is to perform a DNA binding protein pull-down assay using a probe. The following journal article is a good example of how the LightShift Chemiluminescent EMSA Kit and pull-down assays were used to detect a transcription factor bound to a DNA probe: Ragione, A.L., et al. (2003), J. Biol. Chem. 278(26):23360-8.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I am using the LightShift Chemiluminescent EMSA kit. How much protein do I need for each reaction?

The amount of protein extract needed for a binding reaction depends on how much active DNA binding protein is in the sample. The LightShift Kit is sensitive and will easily detect 5 fmol of active protein bound to 5 fmol of biotinylated probe. If the protein being studied is abundant, 0.25 µg of a cell lysate may be sufficient for each binding reaction. However, if the protein of interest is rare, 10 µg or more of cell lysate may be needed. Using a large excess of protein extract may lead to high background signal and non-specific bands.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What is a "supershift"?

A supershift assay is a method for positively identifying a protein:DNA interaction on an EMSA. An antibody (typically 1 µg) is added to the binding reaction. During electrophoresis, the antibody:protein:DNA complex migrates slowly, causing a “supershift” compared to the “shift” caused by a protein:DNA complex. Not all antibodies will cause a supershift. Some antibodies do not bind to proteins once they are bound to DNA. Some antibodies can prevent protein:DNA interactions but can still be used to confirm the identity of a protein that causes a shift in the absence of the antibody.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.