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【产品名称】
 EpiQuik染色质免疫沉淀(ChIP)试剂盒
【订购货号】
 E14527
【产品CAS号码】
 无
【规格/价格】
 
【预计到货时间】
 30 天
【产品详细说明】
【本产品仅供科学研究使用】
【更新日期】 2015-5-26  

产品名称:EpiQuik Chromatin Immunoprecipitation (ChIP) Kit/ EpiQuik染色质免疫沉淀试剂盒(ChIP)

产品货号:P-2002

产品规格:24 reactions,48reactions,96reactions

产品概述

EpiQuik™染色质免疫沉淀(ChIP)试剂盒是一套完整的优化试剂,基于一个方便的微孔板进行染色质免疫沉淀,该试剂盒是即用型并能提供成功完成CHIP实验所需的所有必需试剂。EpiQuik™ ChIP试剂盒适用于特异性免疫沉淀与定量定性PCR、ChIP-Seq和 ChIP-on-chip的结合。

为选择EpiQuik染色质免疫沉淀试剂盒(ChIP)

•快速和简单的微孔板程序,实验能在5小时内完成。

•微孔板的形式使得分析灵活:手动或高通量。

•DNA纯化柱包括在内:节约了大量的时间和减少不必要的体力劳动。

•与所有的DNA扩增方法兼容。

•能达到非常可靠和一致的试验条件。

 

产品详细信息:

该试剂盒包括一个阳性对照抗 体(RNA聚合酶II),阴性对照正常小鼠IgG,和GAPDH引物可作为证明试剂和方案是否有效的阳性对照。在大部分生长中的哺乳动物细胞中RNA聚合酶II会在GAPDH基因启动子上富集,其准备起始转录,因此该启动子能被RNA聚合酶II免疫沉淀,而与正常的小鼠IgG不起作用。

染色质免疫沉淀过程中,用甲醛交联细胞,提取染色质。染色质被剪切并加入到被亲和抗体包被的微孔板中。交联的DNA从抗体-捕获蛋白-DNA复合物中释放出,反转后通过特异设计的快速离心柱纯化。洗脱下来的DNA可用于各种下游应用。

 

EpiQuikTM公司的染色质免疫沉淀试剂盒与其他公司的比较:

 

EpiQuikTM

Other CHIPs

分析时间

小于5小时

3天

高通量

不能

方便(分析步骤)

小于30

大于60

劳动强度

需要细胞数

1-5×105/well

10-50×105/well

PCR反应/DNA洗脱量

20

25-30

P/N信号比例

20-200:1

20-200:1

组织特异性

 

 

Product Overview

The EpiQuik™ Chromatin Immunoprecipitation (ChIP) Kit is a convenient package of tools that allows the experimenter to perform chromatin immunoprecipitation (ChIP) at extraordinarily rapid speeds and consistency, superior to all other current ChIP methods available. The kit is ready-to-use and provides all the essential components needed to carry out a successful ChIP experiment. The EpiQuik™ ChIP kits are suitable for combining the specificity of immunoprecipitation with qualitative and quantitative PCR, MS-PCR, DNA sequencing, and southern blot, as well as DNA microarray.

WHY CHOOSE THE EPIQUIK™ CHROMATIN IMMUNOPRECIPITATION (CHIP) KIT?

  • The number one fastest procedure available. The entire procedure can be completed within an amazingly short 5 hour period with superb results.
  • The ChIP procedure has been drastically simplified -- extremely short and easy to follow.
  • Strip microwell format makes the assay flexible: manual or high throughput.
  • Columns for DNA purification are included: save tremendous amounts of time and reduce unnecessary physical labor.
  • Compatible with all DNA amplification-based approaches.
  • Achieves very reliable and consistent assay conditions.
Product Details


This kit includes a positive control antibody (RNA polymerase II), a negative control normal mouse IgG, and GAPDH primers that can be used as a positive control to demonstrate the efficacy of the kit reagents and protocol. RNA polymerase II is considered to be enriched in the GAPDH gene promoter that is expected to be undergoing transcription in most growing mammalian cells and can be immunoprecipitated by RNA polymerase II but not by normal mouse IgG.

In this ChIP, cells are cross-linked with formaldehyde and chromatin is extracted. The chromatin is then sheared and added into the microwell immobilized with affinity antibodies. Cross-linked DNA is released from antibody-captured protein-DNA complex, reversed and purified through the specifically designed Fast-Spin Column. Eluted DNA can be used for various down-stream applications.

 

 

SCHEMATIC PROCEDURE: COMPARATIVE OVERVIEW:

 

Product Components


CP1 (Wash Buffer)
CP2 (Antibody Buffer)
CP3A (Lysis Buffer)
CP3B (Lysis Buffer)
CP4 (ChIP Dilution Buffer)
CP5 (DNA Release Buffer)
CP6 (Reverse Buffer)
CP7 (Binding Buffer)
CP8 (Elution Buffer)
Protease Inhibitor Cocktail (100X)*
Normal Mouse IgG (1 mg/ml)*
Anti-RNA Polymerase II (1mg/ml)*
Proteinase K (10 mg/ml)*
Control Primer (GAPDH)
Forward (20 µM)*
Reverse (20 µM)*
8-Well Assay Strips (with Frame)
8-Well Strip Caps
F-Spin Column
F-Collection Tube
User Guide

* Spin the solution down to the bottom before use.

Frequently Asked Q's


1. Can IP be overnight?
Yes.

2. Is there any background DNA by mouse IgG IP?
Yes, a little < 2 ng.

3. Can any species Ab can be used for the IP?
Yes, if it is an IgG antibody.

4. Can kit P-2002 be used for frozen tissues?
Yes, but it is not as good as fresh samples.

5. How much volume of elution should be added into PCR reaction?
2 µl.

6. Are cell extracts containing 1%SDS compatible with CP4?
No, the SDS concentration in IP solution should be less than 0.1%.

7. Why is there no difference of CT values between the negative control and samples?
A. Increase washing time by one.
B. Check if antibody is chip grade.
C. There is no enrichment on the promoter.

8. Why is the difference between negative control and positive control not significant?
A. For conventional PCR, reduce PCR cycles to 35-36 or optimize the PCR cycles.
B. Increase washing time by one additional wash.

User Guide & MSDS


[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

[Material Safety Data Sheet]

Product Citations


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