Pierce™ 磁性 ChIP 试剂盒
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Pierce™ 磁性 ChIP 试剂盒

Thermo Scientific Pierce 磁性 ChIP 试剂盒提供了一种通过免疫沉淀(染色质 IP)高效分离染色质结合 DNA了解更多信息
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货号数量
261571 个试剂盒
货号 26157
价格(CNY)
5,375.00
飞享价
Ends: 31-Dec-2025
7,792.00
共减 2,417.00 (31%)
Each
添加至购物车
数量:
1 个试剂盒
请求批量或定制报价
价格(CNY)
5,375.00
飞享价
Ends: 31-Dec-2025
7,792.00
共减 2,417.00 (31%)
Each
添加至购物车
Thermo Scientific Pierce 磁性 ChIP 试剂盒提供了一种通过免疫沉淀(染色质 IP)高效分离染色质结合 DNA 的便利方法,用于随后通过 PCR 进行定量分析。

磁性 ChIP 试剂盒的特点:

快速— 可在约 8 小时内获得适于进行定量 PCR 的纯化 DNA
高效且可重现—微球菌核酸酶酶切过程和细胞核裂解过程均经高度优化
灵敏—只需 10,000 个细胞 (1 x 10^4) 即可获得结果
低背景—Pierce 蛋白 A/G 磁珠已在不含 DNA 的试剂中进行封闭,可极大限度地降低背景
全面—包括优化的阳性对照试剂:RNA 聚合酶 II 抗体和 GAPDH 启动子 PCR 引物

Pierce 磁性染色质免疫沉淀分析 (ChIP) 试剂盒含有足够的试剂,可使用优化的方案采用适当对照品进行 30 次 ChIP 检测。试剂盒中包含的试剂可用于裂解细胞、捕获蛋白-DNA 复合物,逆转交联以及分离 DNA。封闭 Pierce 蛋白 A/G 磁珠具有极高的结合容量和较低的非特异性背景,可与很多种抗体一起使用。这些微珠可与磁力架或自动化平台(例如,Thermo Scientific KingFisher 仪器)一起用于手动工作流程。还提供经验证适用于 ChIP 且具备质量保证的抗体,与 Pierce 磁性 ChIP 试剂盒配合使用。

包括:
试剂盒含有用于染色质制备、IP 以及 DNA 纯化的试剂,同时也包含阳性对照品(抗体和引物)

需要:
体内交联剂(如甲醛)、微量吸头超声波仪(如 Misonix™ 超声波仪 3000)、符合 ChIP 要求的首选抗体、目标 DNA 序列的 PCR 引物、PCR 预混液(如果需要 qPCR,则含有一种染料,如 SYBR™ Green)和 qPCR 仪器

应用:
• 通过 PCR 或 ChIP-Seq 确定基因组 DNA 上特异性蛋白-DNA 相互作用的位点
• 监测组蛋白修饰或化学制剂对 DNA 结合的影响

要想成功进行 ChIP 检测,需要在检测靶标基因组 DNA 之前执行多个关键步骤(交联、染色质制备、免疫沉淀)。单独来讲,ChIP 实验方案中的每一步在优化时都很费时间。Thermo Scientific Pierce 磁性 ChIP 试剂盒简化了 ChIP 流程,可获得准确且可重现的结果。

蛋白-DNA 复合物首先在体内与甲醛交联。试剂盒含有的试剂可以裂解细胞,并可提取和溶解交联复合物。然后,将复合物和特异性抗体放在一起进行孵育,最后用 Pierce 蛋白 A/G 磁珠进行分离。在逆转交联和酶切蛋白后,纯化所得 DNA 片段,然后准备进行标准或定量 PCR。

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相关产品
Pierce™ 磁性 ChIP 试剂盒
Pierce™ ChIP 级蛋白 A/G 磁珠
仅供科研使用。不可用于诊断程序。
规格
描述Pierce 磁性 ChIP 试剂盒
产品规格试剂盒
套装内容4x10^6 个细胞/反应
数量1 个试剂盒
足够用于30 次反应
容量(公制)4×10^6 个细胞/反应
产品线Pierce™
类型磁珠 ChIP 试剂盒
Unit SizeEach
内容与储存
• 甘氨酸溶液 (10X),15 mL;在室温下储存
• PBS (20X),15 mL;在 4°C 下储存
• Halt™ 蛋白酶和磷酸酶抑制剂混合物,无 EDTA (100X),1 mL;在 4°C 下储存
• 膜提取缓冲液,10 mL;在 4°C 下储存
• MNase 酶切缓冲液,10 mL;在 4°C 下储存
• DTT (7.7 mg),冻干,2 个样品瓶;在室温或 4°C 下储存
• MNase 终止液,1 mL;在 4°C 下储存
• ChIP 级蛋白 A/G 磁珠,0.7 mL;在 4°C 下储存
• IP 稀释/洗涤缓冲液 (5X),40 mL,在 4°C 下储存
• 氯化钠 (5M),6 mL;在 4°C 下储存
• IP 洗脱缓冲液 (2X),4.5 mL;在 4°C 下储存
• 微量离心管,1.5 mL,75 个管;在室温或 4°C 下储存
• DNA 纯化柱和试剂,40 次纯化;在室温或 4°C 下储存
• DNA 纯化柱,40 个柱
• DNA 柱结合溶液,30 mL
• DNA 柱洗涤液,6 mL
• pH 指示剂,0.8 mL
• DNA 柱洗脱液,5 mL
• 微球菌核酸酶(ChIP 级)(10 U/µL),50 µL;在-20°C 下储存
• 抗 RNA 聚合酶 II 抗体,25 µL;在-20°C 下储存
• 正常兔 IgG (1 mg/mL),10 µL;在-20°C 下储存
• 蛋白酶 K (20 mg/mL),0.25 mL;在-20°C 下储存
• ChIP 阳性对照引物(GAPDH 启动子,人特异性),100 µL;在 -20°C 下储存

常见问题解答 (FAQ)

With my ChIP assay (Agarose ChIP Kit/Magnetic ChIP Kit), the qPCR signal (cycle threshold) from my positive and negative controls show no difference or are very close. Why is this?

-Verify that your specific antibody (if not using the kit-provided RNA polymerase II antibody) is validated for IP. Ideally, a ChIP validated antibody is the best, but an antibody for IP has a good chance of working in ChIP.
-Ensure that your chromatin is properly digested (see Appendix A in the manual). Too much digestion as well as too little digestion will affect the success of the ChIP reaction.
-Ensure that all the chromatin has been released from the nuclei. When following the Magnetic ChIP kit instructions, MNase digestion of 4x106 cells followed by sonication to lyse the nuclei, yields about 20-50 µg for the IP. This same sequence can be used with the Agarose ChIP Kit as well. It is recommended that you start with 2–4 x 106 cells per ChIP reaction. Once a successful ChIP has been run at this cell number, it is possible to decrease the cell amount empirically. We have seen good results using as little at 10,000 cells, but this entirely depends on the cell line, target, and antibody.
-Ensure that enough DNA was used for qPCR. Typically, 30-80 ng of DNA is a good range.

With my Magnetic ChIP Kit, what can cause no or low PCR signal in the experimental IP samples?

Here are possible causes and solutions:

- Insufficient amount of antibody added to the IP: Add more antibody to the IP.
- Antibody did not function in an IP: Verify that the antibody is qualified for CHIP or IP applications and has been handled and stored properly.

With my Magnetic ChIP Kit, what can cause PCR signal of the positive and negative control IP samples to be equivalent?

Here are possible causes and solutions:

- Excess chromatin or antibody added to the IP: Add less chromatin or antibody.
- PCR amplification was measured outside the linear range of amplification: Decrease the number of amplification cycles used in the PCR reaction.
- Insufficient amount of sample DNA added to the PCR reaction: Add more sample DNA to the PCR reaction.

With my Magnetic ChIP Kit, what can cause no or low PCR signal in the positive control IP samples?

Here are possible causes and solutions:

- Insufficient chromatin amount in the IP reaction: Use at least 25 µg of chromatin for each IP.
- Insufficient antibody incubation time: Incubate antibody overnight.
- Nuclei not fully lysed: Monitor sonication of nuclei by microscope to ensure full lysis.
- Low-abundance target: Add more chromatin or magnetic beads (30 µL).

With my Magnetic ChIP Kit, what can cause no or low PCR signal in the total input control samples?

Here are possible causes and solutions:

- PCR amplification conditions were not fully optimized: Optimize PCR conditions using samples known to contain the target amplicon; Check primer design
- Insufficient amount of sample DNA added to the PCR reaction: Increase the amount of sample DNA added to the PCR reaction
- Nuclei not fully lysed: Monitor sonication of nuclei by microscope to ensure full lysis