成千上万次的ChIP 实验告诉您，这样的结果才是“真阳性”

成人的字典里没有“容易”二字，而做科研的人似乎对“不容易”有更深刻的理解。每做一个新的实验犹如西游记中唐憎取经路上的一次与妖魔鬼怪的搏斗，每一次实验的成功好似孙悟空降服了各路妖怪。作为初学者，一开始做常规的染色质免疫共沉淀（ChIP)实验，得到完好的DNA 需要耗时2天左右，而在这两天的实验过程中，我们要对ChIP 实验的每一步都再三斟酌，对染色质的质量做准确无误的质控后才能开始后续的IP与纯化等步骤。即使实验每一步都能顺利，但最终是否取得了真（zhen）阳性（jing），还需要多个指标来评价。

ChIP实验是通过抗体把DNA-Protein 的复合物沉淀下来，然后对与蛋白结合的 DNA 进行纯化， 随后，进一步对DNA 进行普通PCR, qPCR 或者测序等实验来分析结果。那如何来评价和计算ChIP-PCR的结果呢？

一般情况下，一次严格的ChIP 实验会设置阳性对照组、阴性对照组、目的蛋白组及input对照。我们通常看到普通PCR 的结果是图1这样的，这里研究了PPARγ与LPL （脂蛋白脂肪酶,lipoprotein lipase) 启动子区的结合情况。与染色质稳定结合的RNA poll-II 作为阳性对照，而ICAM-1是一个唯一定位于胞膜的蛋白，可以作为阴性对照。很多时候，阴性对照也可以是与目的抗体同源的IgG。

图1  普通PCR 结果示意图

但是CST推荐使用实时PCR(qPCR)来分析CIP 结果，因为它是定量的。因此，此文主要讲的是ChIP-qPCR 的设置、计算与评价。

关于 PCR 循环数

对于普通的PCR的单拷贝基因，我们建议使用大约30个PCR扩增循环； 对于多拷贝基因，如α卫星重复序列，我们建议使用大约20个PCR扩增循环，以确保PCR反应在线性扩增范围内。如果进行太多循环，则可以观察到Input样品，目的蛋白IP样品和IgG样品之间的差异较小，因为PCR扩增超出线性扩增范围并且所有信号变得饱和。 如果使用实时定量PCR(qPCR)，则推荐40个循环的扩增，因为qPCR仪器能够实时记录每一循环的产物信号，通过还原完整扩增曲线来计算最初模板DNA的丰度。

ChIP-qPCR 的设置

ChIP-qPCR 的计算

计算CHIP-qPCR 富集效率时，一般是以相对Input信号富集比例来呈现的，例如若做了单个2%Input,计算公式如下：

注：CT 值一般是指2-3次重复实验的平均CT值

而 CST 一直推荐通过稀释不同比例的Input 来先建立qPCR  的标准曲线，标准曲线一般用来确定qPCR的反应效率。低扩增效率（小于90%）可能由于Taq酶抑制剂的污染、过高或未优化的退火温度、时间较久或已失活的Taq酶、引物设计不合理或扩增子含二级结构等。过高反应率（大于105%）一般是由于引物二聚体或者非特异性扩增。我们建议把Input 设置4个依次5倍稀释的点来确保标准曲线能覆盖实验中所有可能会用到的模版浓度（即形成宽的动态范围）。以总input染色质的2%开始，我们用这个稀释梯度系列(2%、0.4%、0.08%、0.016%)来为每个input染色质qPCR样品建CT的标准曲线。标准曲线是模版起始浓度的对数值对CT 值得到的，其后所得的线性回归直线方程，Pearson 相关系数（r）或决定系数（R2）可用来评价qPCR 反应是否需要优化。以下是如何制作标准曲线的方法及计算PCR 扩增效率的公式，可供参考。

注：Slope：标准曲线的斜率；Y intercept：标准曲线截距

这样我们就能通过将每个免疫沉淀样品的qPCR反应测出来的CT值转变为表示总input染色质的百分比的值，来计算每个免疫沉淀样品的DNA丰富度。

ChIP-qPCR 的评价

qPCR结果计算出来以后，我们还需要进一步比较阳性、阴性对照相对input富集丰度及目的信号相对于阳性、阴性对照富集丰度的倍数。经过成千上万次的ChIP-qPCR，CST 科学家给出了Q-PCR标准质控：

1）组蛋白H3阳性对照相对input富集丰度正常值在1%-5%左右；

2）阴性对照IgG相对input富集丰度应低于0.1%。

3）当靶标抗体在目标基因位点的相对富集丰度，相比于非目标基因位点（或阴性对照引物）的富集丰度比值超过3倍，相对于阴性对照IgG的相对富集丰度比值达到8到10倍以上时，ChIP结果信噪比达到合理范围，我们定义ChIP结果为阳性。

4）一般情况下，组蛋白相对于input富集丰度在1-50%范围；转录因子或辅因子相对于input富集丰度在0.2-1%。

具体如下：

真阳性的结果往往是图2这样的：

      图2  真阳性ChIP–qPCR 结果

而假阳性的结果是图3这样的，主要的原因是IP 下来的DNA没有或不足 、PCR条件不合理等。

 图3  忽悠人的假阳性结果

如果想了解更多的ChIP 相关知识，可参考文末「猜你喜欢」。

请不要被“假阳性”蒙蔽了双眼。吃一堑，长一智。成千上万次的实验经验造就了金睛火眼，ChIP-qPCR唯有按CST 科学家严格的质控标准，才能检验出“真阳性”。

最后，CST 为所有ChIP验证的抗体提供阳性和阴性对照引物组，可点击此处查询，或直接翻看下方表格。

  需滑动以看完整表格

No.	Antibody	Reactivity	Positive Control Primers	Negative Control Primers
14129	AEBP2 (D7C6X) Rabbit mAb	H, M, R, Mk, (Hm, C, B)	7707 H, 5517 H	4486 H
83200	AhR (D5S6H) Rabbit mAb	H	81126 H	4486 H
15103	Aiolos (D1C1E) Rabbit mAb	H, M	12899 H	4486 H
8486	RUNX2 (D1H7) Rabbit mAb	H, M, R, Mk	12376 H	4486 H
12556	RUNX2 (D1L7F) Rabbit mAb	H, M, R	12376 H	4486 H
33593	ATF-3 (D2Y5W) Rabbit mAb	H, M	66427 H	4486 H
11815	ATF-4 (D4B8) Rabbit mAb	H, M, R	13059 M	7015 M
14820	ATRX (D1N2E) Rabbit mAb	H	14901 H	5516 H
80775	BACH2 (D3T3G) Rabbit mAb	H	53538 H	4486 H
12120	Bcl-11B (D6F1) XP® Rabbit mAb	H, (M, R, Mk)	11951 H	4486 H
14895	BCL6 (D4I2V) XP® Rabbit mAb	H	12899 H	4486 H
5650	BCL6 (D65C10) Rabbit mAb	H, M	12924 H, 12899 H	4486 H
14020	BMAL1 (D2L7G) Rabbit mAb	H, M, R	13902 M	7015 M
6964	Bmi1 (D20B7) XP® Rabbit mAb	H, Mk	7707 H, 5517 H	4486 H
5856	Bmi1 (D42B3) Rabbit mAb	H, M, R, Mk	7707 H, 5517 H	4486 H
5848	Brd2 (D89B4) Rabbit mAb	H, M, (R, Mk)	62858 H	4486 H
13440	BRD4 (E2A7X) Rabbit mAb	H, (B, Dg, Pg)	12924 H	4486 H
11974	RNF20 (D6E10) XP® Rabbit mAb	H, M, R, Mk, (Hm, Dg, Pg, GP,  Hr)	5037 H, 4471 H	4486 H
49360	Brg1 (D1Q7F) Rabbit mAb	H, M, R, Mk	9673 H, 9702 H	4486 H
11966	BRM (D9E8B) XP® Rabbit mAb	H, Mk, (Dg)	9673 H, 9702 H	4486 H
8814	Non-phospho (Active) β-Catenin (Ser33/37/Thr41)  (D13A1) Rabbit mAb	H, M, R, Mk, (C, X, Z, B, Dg,  Pg, GP, Hr)	8973 H, 5111 H	4486 H
8480	β-Catenin (D10A8) XP® Rabbit mAb	H, M, R, Mk, (Z, B, Pg, GP, Hr)	8973 H, 5111 H	4486 H
9581	β-Catenin Antibody (Amino-terminal Antigen)	H, M, R, Mk	5111 H	4486 H
9587	β-Catenin Antibody (Carboxy-terminal Antigen)	H, M, R, Mk, (C, X, B, Dg, Pg,  Hr)	8973 H, 5111 H	4486 H
62184	CBFβ (D4N2N) Rabbit mAb	H, M, R, Mk	29965 M	8986 M
4771	Acetyl-CBP (Lys1535)/p300 (Lys1499) Antibody	H, M, Mk, (R)	4829 H	4486 H
7389	CBP (D6C5) Rabbit mAb	H, M, R, Mk	4829 H	4486 H
7425	CBP (D9B6) Rabbit mAb	H, M, R, Mk	4829 H	4486 H
14696	CBX8 (D2O8C) Rabbit mAb	H, M, R, Mk, (Hm, B)	7707 H, 5517 H	4486 H
81464	Cyclin T1 (D1B6G) Rabbit mAb	H, M, R, Mk	5516 H, 14905 H	4486 H
4351	CHD1 (D8C2) Rabbit mAb	H, M, R, Mk, (B, Dg, Pg)	4653 M, 7015 M	8985 M
12011	CHD4 (D4B7) Rabbit mAb	H, M, R, Mk, (Hm, B, Pg, GP,  Hr)	5516 H, 7014 H	4490 H
2895	CHOP (L63F7) Mouse mAb	H, M, R	13059 M	7015 M
14894	COBRA1 (D6K9A) Rabbit mAb	H, M, R, Mk, (Hm, B)	14905 H	4486 H
9198	Phospho-CREB (Ser133) (87G3) Rabbit mAb	H, M, R, (Z)	4829 H	4486 H
9197	CREB (48H2) Rabbit mAb	H, M, R, Mk, Dm	4829 H	4486 H
4820	CREB (D76D11) Rabbit mAb	H, M, R, Hm, Mk, Dm	4829 H	4486 H
3417	CTCF (D1A7) XP® Rabbit mAb	H, R, Mk, (B)	5172 H	4486 H
3418	CTCF (D31H2) XP® Rabbit mAb	H, M, R, Mk, (B, Hr)	5172 H	4486 H
2899	CTCF Antibody	H, M, R, Mk	5172 H	4486 H
3742	E2F-1 Antibody	H, (R, B)	7001 H	4486 H
14159	EAF2 (E1R8C) Rabbit mAb	H	14023 H	4486 H
4153	EGR1 (15F7) Rabbit mAb	H, M, R, (B)	5549 H	4486 H
4154	EGR1 (44D5) Rabbit mAb	H, M, R, (B)	5549 H	4486 H
14468	ELL (D7N6U) Rabbit mAb	H, M, R, (B, Dg, Hr)	14084 H	4486 H
13258	Estrogen Receptor α (D6R2W) Rabbit mAb	H	9673 H, 9702 H	4486 H
8644	Estrogen Receptor α (D8H8) Rabbit mAb	H	9702 H	9673 H
13826	ERRα (E1G1J) Rabbit mAb	H, M, R, Mk, (B, Dg, Pg, Hr)	13828 M	8985 M
14069	ETS-1 (D8O8A) Rabbit mAb	H, M, R	4663 H, 14905 H	4486 H
5246	Ezh2 (D2C9) XP® Rabbit mAb	H, M, R, Mk	7707 H, 5517 H	4486 H
4905	Ezh2 Antibody	H, M, R, Pg	7707 H, 5517 H	4486 H
5348	Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb	H, M, R, (Hm, Mk, B, Pg, Hr)	7983 R	7964 R
2250	c-Fos (9F6) Rabbit mAb	H, M, R, (Hm, B, Pg)	7983 R	7964 R
2251	FosB (5G4) Rabbit mAb	H, M, R	7983 R	7964 R
4402	FoxP1 (D35D10) XP® Rabbit mAb	H, M, R, Mk	12924 H, 12899 H	4486 H
5841	Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb	H, M, R, (Mk, B, Hr)	7983 R	7964 R
19967	Fra2 (D2F1E) Rabbit mAb	H, (B, Hr)	7983 R	7964 R
3306	G9a/EHMT2 (C6H3) Rabbit mAb	H, M, R, Mk, (B, Pg, Hr)	98428 H	5516 H
5852	GATA-3 (D13C9) XP® Rabbit mAb	H, M, (Mk)	13051 H	4486 H
5851	GATA-6 (D61E4) XP® Rabbit mAb	H, (M, R, Dg, Pg)	13840 H	4486 H
12041	Glucocorticoid Receptor (D6H2L) XP® Rabbit  mAb	H, M, R, Mk	7681 H	4486 H
3660	Glucocorticoid Receptor (D8H2) XP® Rabbit mAb	H, M, R, Mk	7681 H	4486 H
41328	Histone H1.4 (D4J5Q) Rabbit mAb	H, Mk, (Hm, B, Dg)	4486 H, 7014 H	5037 H, 5516H
75336	Acetyl-Histone H2AZ (Lys4/Lys7) (D3V1I) Rabbit mAb	H, M, R	62086 H	4486 H
3724	HA-Tag (C29F4) Rabbit mAb	all species expected
34589	HDAC1 (D5C6U) XP® Rabbit mAb	H, M, R, Mk	5516 H, 7014 H	95064 H
57156	HDAC2 (D6S5P) Rabbit mAb	H, M, R, Mk	7014 H	95064 H
85057	HDAC3 (D2O1K) Rabbit mAb	H, M, R, Mk	9673 H, 9702 H	4486 H
42427	Helios (D8W4X) XP® Rabbit mAb	H, M	99663 H	4486 H
36426	Helios (D4Z6D) Rabbit mAb	H	99663 H	4486 H
5537	HIF-1β/ARNT (D28F3) XP® Rabbit mAb	H, M, R, Mk	81126 H	4486 H
14179	HIF-1α (D2U3T) Rabbit mAb	H, M, R, Mk	75671 H	4486 H
8240	Ubiquityl-Histone H2A (Lys119) (D27C4)  XP® Rabbit mAb	H, M, R, Mk	7707 H, 5517 H	4486 H, 5516H
12349	Histone H2A (D6O3A) Rabbit mAb	H, M, R, Mk, Z, (Hm, B, Dg, GP)	4486 H, 5516 H, 4493 H, 7014 H
12799	Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit  mAb	H, M, R, Mk, (Hm, C, Z, B, Hr)	5516 H, 7014 H	4486 H, 4490H
9072	Acetyl-Histone H2B (Lys12) (D7H4) Rabbit mAb (ChIP  Formulated)	H, (M, R, Mk, C, X, Z, B, Pg,  Hr)	5516 H, 7014 H
9083	Acetyl-Histone H2B (Lys15) (D8H1) XP® Rabbit  mAb	H, M, R, Mk, (Z, B, Pg, Hr)	7014 H	4486 H
5546	Ubiquityl-Histone H2B (Lys120) (D11)  XP® Rabbit mAb	H, M, R, Mk	5047 H, 4478 H	5037 H, 4471H
12364	Histone H2B (D2H6) Rabbit mAb	H, M, R, Mk, (C, Z, B)	4486 H, 5516 H, 4493 H, 7014 H
13576	Phospho-Histone H3 (Thr3) (D5G1I) Rabbit mAb	H, M, (R, Z, Sc)	4486 H	5516 H
53348	Phospho-Histone H3 (Ser10) (D7N8E) XP® Rabbit  mAb	H, M, R, Mk, (Hm, X, Sc)	5037 H, 4471 H
5326	Mono-Methyl-Histone H3 (Lys4) (D1A9)  XP® Rabbit mAb	H, M, R, Mk, (Dm)	5047 H	5037 H
4473	Pan-Methyl-Histone H3 (Lys9) (D54) XP® Rabbit  mAb	H, M, R, Mk, (C, Dm, X, Z, B,  Pg, Sc, Ce)	5098 H, 4486 H, 5077 H	4471 H, 7014H
14186	Mono-Methyl-Histone H3 (Lys9) (D1P5R) Rabbit mAb	H, M, R, Mk, (X, Z, Pg, Sc, Hr)	4478 H, 4490 H	4471 H
14111	Mono-Methyl-Histone H3 (Lys36) (D9J1D) Rabbit mAb	H, M, R, Mk, (Hm, B)	5516 H, 4490 H, 4493 H	5098 H
12522	Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb	H, M, R, Mk	4478 H	4486 H
9649	Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb	H, M, R, Mk, Z, Ce, (Sc)	5516 H, 7014 H, 7015 M	4486 H, 4490H
9677	Acetyl-Histone H3 (Lys9/Lys14) Antibody	H, M, R, Mk, Ce, (Z)	7014 H, 7015 M	4486 H, 4490H
7627	Acetyl-Histone H3 (Lys14) (D4B9) Rabbit mAb	H, M, R, Mk, (Hm, Dm, X, Z, Pg,  Sc, Hr)	7014 H, 7015 M	4486 H, 4490H
13998	Acetyl-Histone H3 (Lys18) (D8Z5H) Rabbit mAb	H, M, R, Mk, Sc, (M, Hm, Pg)	5516 H, 7014 H	4486 H, 4490H
9675	Acetyl-Histone H3 (Lys18) Antibody	H, M, R	5516 H, 7014 H, 7015 M	4486 H, 4490H
8173	Acetyl-Histone H3 (Lys27) (D5E4) XP® Rabbit  mAb	H, M, R, Mk, (Hm, X, Z, GP, Hr)	5516 H, 7014 H, 7015 M	5098 H, 4486H
4353	Acetyl-Histone H3 (Lys27) Antibody	H, M, R, Mk, (Hm, C, X, Z, B)	5516 H, 7014 H, 7015 M	5098 H, 4486H
27683	Acetyl-Histone H3 (Lys36) (D9T5Q) Rabbit mAb	H, M, R, Mk, (Hm, Dm, X, Z, Pg)	13653 H, 7014 H	4486 H, 4493H
9725	Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb	H, M, R, Mk	7014 H, 7015 M	4486 H, 4490H
4658	Di-Methyl-Histone H3 (Lys9) (D85B4)  XP® Rabbit mAb	H, M, R, Mk, (Dm, X, Z, B, Pg,  Sc, Ce)	5098 H, 4486 H	4471 H, 7014H
9753	Di-Methyl-Histone H3 (Lys9) Antibody	H, M, R, Mk, Dm	5098 H, 4486 H	5516 H, 7014H
9728	Di-Methyl-Histone H3 (Lys27) (D18C8)  XP® Rabbit mAb	H, M, R, Mk	5098 H, 4490 H	5516 H, 7014H
5427	Di-Methyl-Histone H3 (Lys79) (D15E8)  XP® Rabbit mAb	H, M, R, Mk	5047 H	5037 H
9751	Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb	H, M, R, Mk, Dm, Sc, (X, Z)	5516 H, 7014 H, 7015 M	4486 H, 4490H
9727	Tri-Methyl-Histone H3 (Lys4) Antibody	H, M, R, Mk, (X, Z)	5516 H, 7014 H, 7015 M	4486 H, 4490H
13969	Tri-Methyl-Histone H3 (Lys9) (D4W1U) Rabbit mAb	H, M, R, Mk, (B)	4486 H	5516 H
5327	Di/Tri-Methyl-Histone H3 (Lys9) (6F12) Mouse mAb	H, M, R, Mk	5098 H, 4486 H	5516 H
9733	Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb	H, M, R, Mk, (X, Z)	4490 H, 4493 H	7014 H
4909	Tri-Methyl-Histone H3 (Lys36) (D5A7)  XP® Rabbit mAb	H, M, R, Mk, (Hm, C, Dm, X, Z,  B)	5047 H, 4478 H	5037 H, 4471H
4620	Histone H3 (D2B12) XP® Rabbit mAb (ChIP  Formulated)	H, M, (R, Hm, Mk, C, Dm, X, Z,  B)	4486 H, 7964 R, 7014 H, 7015 M
14269	Histone H3 (1B1B2) Mouse mAb	H, M, R, Mk	4486 H, 5516 H, 8986 M, 4493 H, 7014 H, 7015 M
2650	Histone H3 Antibody (ChIP Formulated)	H, M, (R, Mk, C, Dm, X, Z, B,  Hr)	4486 H, 7014 H, 7015 M
8647	Acetyl-Histone H4 (Lys5) (D12B3) Rabbit mAb	H, M, R, Mk, (C, Dm, X, Z, B,  Pg, Hr, Ce)	7014 H	4486 H
9672	Acetyl-Histone H4 (Lys5) Antibody	H, M, R, Mk, (C, Dm, X, Z, B,  Pg, Hr, Ce)	7014 H	4486 H
13944	Acetyl-Histone H4 (Lys12) (D2W6O) Rabbit mAb	H, R, Mk, (C, Dm)	5516 H, 7014 H	4486 H
13534	Acetyl-Histone H4 (Lys16) (E2B8W) Rabbit mAb	H, M, R, Mk	5516 H, 7014 H
5737	Tri-Methyl-Histone H4 (Lys20) (D84D2) Rabbit mAb	H, M, R, Mk, (X, B, Pg)	5098 H, 4486 H	5516 H
14149	Histone H4 (D2X4V) Rabbit mAb (ChIP Formulated)	H, (M, R, Hm, Mk, C, B)	4486 H, 4493 H
2616	HP1α Antibody	H, M, R, Mk, (B)	12870 M	12928 M
8676	HP1β (D2F2) XP® Rabbit mAb	H, M, R, Mk, (Hm, B, GP)	12870 M	12928 M
12972	HSF1 (D3L8I) Rabbit mAb	H, M, R, Mk, B, Dg, Pg	5551 H	4486 H
4356	HSF1 Antibody	H, M, R, Mk	5551 H	4486 H
9034	Ikaros (D10E5) Rabbit mAb	H	7273 H	4486 H
14859	Ikaros (D6N9Y) Rabbit mAb	H, M	7273 H	4486 H
76041	IκB-ζ (D4I7C) Rabbit mAb (ChIP Preferred)	M	39609 M	8985 M
13945	INTS9 Antibody	H, M, R, Mk, (Hm, B, Dg, Pg)	14084 H	4486 H
4299	IRF-4 (D43H10) Rabbit mAb	H, R	5156 H	4486 H
4964	IRF-4 Antibody	H	5156 H	4486 H
5628	IRF-8 (D20D8) Rabbit mAb	H, M, (R, Mk, X, B)	9013 H	4486 H
76684	IRF-9 (D2T8M) Rabbit mAb	H	57949 H	4486 H
15106	IRF-4 (D9P5H) Rabbit mAb	H, M, R	5156 H	4486 H
13594	JARID2 (D6M9X) Rabbit mAb	H, M, (Hr)	7707 H, 5517 H	4486 H
3270	Phospho-c-Jun (Ser73) (D47G9) XP® Rabbit mAb	H, M, R, Mk, Pg, (B)	7983 R	7964 R
9165	c-Jun (60A8) Rabbit mAb	H, M, R, Mk	7983 R	7964 R
3753	JunB (C37F9) Rabbit mAb	H, M, R, Mk	7983 R	7964 R
76010	LEF1 (D6J2W) Rabbit mAb	H	8973 H	4486 H
2184	LSD1 (C69G12) Rabbit mAb	H, M, R, Mk	14271 H	4486 H
2139	LSD1 Antibody	H, M, R, Mk	14271 H	4486 H
9814	Acetylated-Lysine (Ac-K2-100) MultiMab™ Rabbit mAb  mix	all species expected	4471 H, 5516 H	4486 H, 4493H
9441	Acetylated-Lysine Antibody	all species expected	5516 H, 7014 H, 7015 M	4490 H, 4493H
11959	MAML1 (D3E9) Rabbit mAb	H, M, R, Mk	7273 H	4486 H
12166	MAML1 (D3K7B) Rabbit mAb	H, M, R	7273 H	4486 H
99169	MBD3 (D1B8F) Rabbit mAb	H, M, R, Mk	5516 H	95064 H
3619	MCM2 (D7G11) XP® Rabbit mAb	H, M, R, Mk	5148 H	4486 H
14950	MED26 (D4B1X) Rabbit mAb	H	14023 H, 14905 H	4486 H
97800	MITF (D3B4T) Rabbit mAb	H, M, Mk	12970 H	4486 H
12590	MITF (D5G7V) Rabbit mAb	H, M, R, Hm, Mk	12970 H	4486 H
14893	MLLT1/ENL (D9M4B) Rabbit mAb	H	14909 H	4486 H
85570	MLX (D8G6W) Rabbit mAb	H, M, R, Mk	95288 M	53559 M
5415	Mouse (G3A1) mAb IgG1 Isotype Control
5646	MTA1 (D17G10) Rabbit mAb	H, M, R, Mk	7014 H	95064 H
13987	c-Myc (D3N8F) Rabbit mAb	H, M, R, Mk	4779 H	4486 H
9402	c-Myc Antibody	H, M, R, (Pg)	4779 H	4486 H
84406	N-Myc (D1V2A) Rabbit mAb	H, M	90678 H	4486 H
51705	N-Myc (D4B2Y) Rabbit mAb	H, M, (R)	90678 H	4486 H
58418	MYST2 (D4N3F) Rabbit mAb	H, M, R, Mk	7707 H, 14909 H	4486 H
8822	Nanog (D2A3) XP® Rabbit mAb (Mouse Specific)	M	4653 M, 4659 M	7015 M
5232	Nanog (D73G4) XP® Rabbit mAb (ChIP  Formulated)	H	4641 H, 4649 H	4486 H
8785	Nanog (D1G10) Rabbit mAb (Mouse Specific; ChIP  Formulated)	M	4653 M, 4659 M	7015 M
3580	Nanog Antibody	H	4641 H, 4649 H	4486 H
4373	NeuroD (D35G2) Rabbit mAb	H, M, R	5549 H	4486 H
12540	NF-κB1 p105/p50 (D7H5M) Rabbit mAb	H, M, (Mk)	5552 H	4486 H
3035	NF-κB1 p105/p50 Antibody	H, Mk	5552 H	4486 H
13586	NF-κB1 p105/p50 (D4P4D) Rabbit mAb	H, M, R	5552 H	4486 H
8242	NF-κB p65 (D14E12) XP® Rabbit mAb	H, M, R, Hm, Mk, Dg	5552 H	4486 H
6956	NF-κB p65 (L8F6) Mouse mAb	H, M, R, Hm, Mk, Mi, B, Dg, Pg	5552 H	4486 H
37359	NF-κB2 p100/p52 (D7A9K) Rabbit mAb	H	5552 H	4486 H
3608	Notch1 (D1E11) XP® Rabbit mAb	H, M, R	7273 H	4486 H
4147	Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb	H, M, R	7273 H	4486 H
46743	NRF1 (D9K6P) Rabbit mAb	H, M, R, Mk	26727 H	4486 H
12721	NRF2 (D1Z9C) XP® Rabbit mAb	H, M, Mk	12635 M	7015 M
8157	Oct-1 (D7B6) Rabbit mAb	H	9673 H, 9702 H	4486 H
5677	Oct-4A (C30A3C1) Rabbit mAb (ChIP Formulated)	H, M	4641 H, 4653 M, 4649 H, 4659 M	4486 H, 7015M
2890	Oct-4A (C52G3) Rabbit mAb	H	4641 H, 4649 H	4486 H
2750	Oct-4 Antibody	H, (Mk)	4641 H, 4649 H	4486 H
83932	Oct-4A (D6C8T) Rabbit mAb (Mouse Specific)	M	4653 M, 4659 M	7015 M
9284	Phospho-p53 (Ser15) Antibody	H, M, R, Mk, (Mi, B, Pg)	6449 H	4486 H
2527	p53 (7F5) Rabbit mAb	H, Mk	6449 H	4486 H
32532	p53 (D2H9O) Rabbit mAb (Rodent Specific)	M, R	76144 M	7015 M
2524	p53 (1C12) Mouse mAb	H, M, R, Hm, Mk	6449 H	4486 H
48818	p53 (DO-7) Mouse mAb	H	6449 H	4486 H
9282	p53 Antibody	H, Mk	6449 H	4486 H
13109	p63-α (D2K8X) XP® Rabbit mAb	H	6449 H	4486 H
59019	PAX8 (D2S2I) Rabbit mAb	H, Mk, (R)	14927 H	4486 H
3378	PCAF (C14G9) Rabbit mAb	H, M, R, Mk, (B, Hr)	4829 H	4486 H
13768	PHC1 (1F3F3) Mouse mAb	H, M	7707 H, 5517 H	4486 H
24799	POLR1A (D6S6S) Rabbit mAb	H, M, Mk	14901 H	13653 H, 4486H
13499	Phospho-Rpb1 CTD (Ser2) (E1Z3G) Rabbit mAb	H, M, R, Mk, (Hm, Dm, X, Z, B,  Pg, Sc, Ce)	13653 H, 13669 H	4486 H
13546	Phospho-Rpb1 CTD (Ser2/Ser5) (D1G3K) Rabbit mAb	H, M, R, Mk, (Hm, Dm, X, Z, B,  Pg, Sc, Ce)	13653 H, 13669 H	4486 H
26319	Phospho-Rpb1 CTD (Thr4) (D7L9W) Rabbit mAb	H, M, R, Mk	20508 H	4486 H
13523	Phospho-Rpb1 CTD (Ser5) (D9N5I) Rabbit mAb	H, M, R, Mk, (Hm, Dm, X, B, Pg,  Sc, Ce)	13653 H, 13669 H	4486 H
13780	Phospho-Rpb1 CTD (Ser7) (E2B6W) Rabbit mAb	H, M, R, Mk	13669 H	4486 H
14958	Rpb1 NTD (D8L4Y) Rabbit mAb	H, M, R, Mk, (Hm, B)	5037 H, 5516 H	5098 H, 4486H
2629	Rpb1 CTD (4H8) Mouse mAb	H, M, R, Mk, (Hm, Dm, Sc)	4471 H, 5516 H	4490 H, 4493H
12825	POLR3A (D5Y2D) Rabbit mAb	H, M, R, Mk, (B, Dg, GP)	12769 H	4486 H
12137	Brn2/POU3F2 (D2C1L) Rabbit mAb	H, M, R	12377 H	4486 H
12495	PRMT4/CARM1 (3H2) Mouse mAb	H, M, R, Mk	12531 H, 9673 H, 9702 H	4486 H
8757	Progesterone Receptor A/B (D8Q2J) XP® Rabbit  mAb	H, (Mk)	7859 H	4486 H
2258	PU.1 (9G7) Rabbit mAb	H, M, (Pg)		4486 H
2266	PU.1 Antibody	H, M, (Mk, Pg)		4486 H
3900	Rabbit (DA1E) mAb IgG XP® Isotype Control
2729	Normal Rabbit IgG
9313	Rb (D20) Rabbit mAb	H, M, Mk	7001 H	4486 H
9309	Rb (4H1) Mouse mAb	H, Mk, B, Pg	7001 H	4486 H
13610	RBL2 (D9T7M) Rabbit mAb	H, M, R, Mk	13707 M	7015 M
13171	RBBP5 (D3I6P) Rabbit mAb	H, M, R, Mk, (C, X, B, Hr)	11951 H	4486 H
5313	RBPSUH (D10A4) XP® Rabbit mAb	H, M, R, Mk	7273 H	4486 H
12659	c-Rel (D5G1A) Rabbit mAb (ChIP Formulated)	H, (R, Mk)	12281 H	4486 H
10544	RelB (D7D7W) Rabbit mAb	H, M, R	5552 H	4486 H
13418	Rev-Erbα (E1Y6D) Rabbit mAb	H, M, R	13413 H	4486 H
5694	RING1B (D22F2) XP® Rabbit mAb	H, M, R, Mk	7707 H, 5517 H	4486 H
74775	Pontin/RUVBL1 (D1L8J) XP® Rabbit mAb	H, M, R, Mk	48781 H	4486 H
61702	SET1A (D3V9S) Rabbit mAb	H	13653 H, 4471 H	4486 H
7691	SIN3A (D1B7) Rabbit mAb	H, M, R, Mk, (Pg, Hr)	9673 H, 5516 H	4486 H, 4490H
8056	SIN3A (D9D6) Rabbit mAb	H, Mk	9673 H, 5516 H	4486 H, 4490H
6944	Smad1 (D59D7) XP® Rabbit mAb	H, M, Mk, (X, B)	5139 H	4486 H
9743	Smad1 Antibody	H, M, Mk	5139 H	4486 H
11971	Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/  Smad9 (Ser465/467) (D5B10) Rabbit mAb (ChIP formulated)	H, (M, R, Mk)	5139 H	4486 H
5339	Smad2 (D43B4) XP® Rabbit mAb	H, M, R, Mk	5139 H, 4669 H	4486 H
8685	Smad2/3 (D7G7) XP® Rabbit mAb	H, M, R, Mk	5139 H, 4669 H	4486 H
5678	Smad2/3 Antibody	H, M, R, Mk, (X)	5139 H, 4669 H	4486 H
9520	Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb	H, M, R, (Mk, X, Z, B)	5139 H, 4669 H	4486 H
9523	Smad3 (C67H9) Rabbit mAb	H, M, R, Mk, (X, Z, B)	5139 H, 4669 H	4486 H
38454	Smad4 (D3M6U) Rabbit mAb	H, M, R, Mk, (Dg, GP, Hr)	5139 H	4486 H
12534	Smad5 (D4G2) Rabbit mAb	H, M, R, Mk	5139 H	4486 H
91735	SMARCB1/BAF47 (D8M1X) Rabbit mAb	H, M, R, Mk	9673 H, 9702 H	4486 H
11956	SMARCC1/BAF155 (D7F8S) Rabbit mAb	H, M, R, Mk	9673 H, 9702 H	4486 H
12760	SMARCC2/BAF170 (D8O9V) Rabbit mAb	H, M, R, Mk, (Dg, Pg, GP, Hr)	9673 H, 9702 H	4486 H
5024	Sox2 (D6D9) XP® Rabbit mAb (ChIP Formulated)	H, (Mk, B, Dg, Hr)	4641 H, 4649 H	4486 H
23064	Sox2 (D9B8N) Rabbit mAb	H, M	4653 M, 4659 M	7015 M
2748	Sox2 Antibody	H, M, (R, Mk, B, Dg, Hr)	4641 H, 4653 M, 4649 H, 4659 M	4486 H, 7015M
9389	SP1 (D4C3) Rabbit mAb	H, Mk	7531 H	4486 H
12191	SPT16 (D7I2K) Rabbit mAb	H, M, R, Mk, (Hm, X, Z, B, Dg,  GP, Hr)	11953 H, 12010 H	5549 H, 4663H
15616	SPT6 (D6J9H) Rabbit mAb	H, M, R, Mk	5516 H, 14905 H	4486 H
2191	SRC-1 (128E7) Rabbit mAb	H, M, R, Mk	25661 H	95064 H
20301	SRC-1 (D1M3Y) Rabbit mAb	H, M, R, Mk	25661 H	95064 H
2126	SRC-3 (5E11) Rabbit mAb	H, M, R, Mk	12531 H	4486 H
5147	SRF (D71A9) XP® Rabbit mAb	H, M, R, Mk, Pg	4663 H	4486 H
21792	SS18 (D6I4Z) Rabbit mAb	H, M, R, Mk, (Dg, Hr)	9673 H, 9702 H	4486 H
13421	SSRP1 (E1Y8D) Rabbit mAb	H, M, R, Mk, (C)	11953 H, 12010 H, 4663 H	5549 H
9167	Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb	H, M	5148 H	4486 H
7649	Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb	H, M, R, (Mk)	5148 H	4486 H
8826	Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb	H, M, R, Mk, (B)	5148 H	4486 H
9177	Phospho-Stat1 (Ser727) Antibody	H, M, R, Mk, (Mk, B)	5148 H	4486 H
14994	Stat1 (D1K9Y) Rabbit mAb	H, M, R, Mk	5148 H	4486 H
14995	Stat1 (D4Y6Z) Rabbit mAb	H, M, R	5148 H	4486 H
9172	Stat1 Antibody	H, M, R, Mk, (B, Dg)	5148 H	4486 H
72604	Stat2 (D9J7L) Rabbit mAb	H, M	30101 H	4486 H
9145	Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb	H, M, R, Mk, (Hm, B, Pg, Hr)	4663 H	4486 H
9131	Phospho-Stat3 (Tyr705) Antibody	H, M, R, Mk, (C, B, Dg)	4663 H	4486 H
34911	Phospho-Stat3 (Ser727) (D4X3C) Rabbit mAb (ChIP  Specific)	H, (M, R)	4663 H	4486 H
9134	Phospho-Stat3 (Ser727) Antibody	H, M, R, (Hm, B, Dg, Pg, Hr)	4663 H	4486 H
8768	Stat3α (D1A5) XP® Rabbit mAb	H, M, R, Hm, Mk, (Pg)	4663 H	4486 H
4904	Stat3 (79D7) Rabbit mAb	H, M, R, Mk	4663 H	4486 H
12640	Stat3 (D3Z2G) Rabbit mAb	H, M, R, Mk, (B, Pg)	4663 H	4486 H
9139	Stat3 (124H6) Mouse mAb	H, M, R, Mk	4663 H	4486 H
4134	Phospho-Stat4 (Tyr693) (D2E4) Rabbit mAb	H, (M, R, Mk, Pg)	9014 H	4486 H
5267	Phospho-Stat4 (Tyr693) Antibody	H, (M, R)	9014 H	4486 H
2653	Stat4 (C46B10) Rabbit mAb	H, M, R	9014 H	4486 H
9351	Phospho-Stat5 (Tyr694) Antibody	H, M, (R, B)	5131 M	7015 M
94205	Stat5 (D2O6Y) Rabbit mAb	H, M, R	5131 M	7015 M
9363	Stat5 Antibody	H, M, R, Mk	5131 M	7015 M
5397	Stat6 (D3H4) Rabbit mAb	H, M, R, (Mk, B, Dg, Pg)	7710 H	4486 H
12800	STF-1 (D1Z2A) XP® Rabbit mAb	H, M, R, (B)	12864 H	4486 H
64828	SPT4 (D3P2W) Rabbit mAb	H, M, R, Mk, (Hm, Sc)	5516 H, 14905 H	4486 H
3737	SUZ12 (D39F6) XP® Rabbit mAb	H, M, R, Mk, (Pg, Hr)	7707 H, 5517 H	4486 H
4883	TAZ (V386) Antibody	H, M, R, (Hr)	14927 H	14928 H
74499	TBL1XR1/TBLR1 (D4J9C) Rabbit mAb	H, Mk	8973 H, 5111 H	4486 H
44059	TBP (D5C9H) XP® Rabbit mAb	H, M, R, Mk, (C, B)	5037 H, 4471 H	4486 H, 4493H
12578	TBP (D5G7Y) Rabbit mAb	H, Mk	5037 H, 4471 H	4486 H
13232	T-bet/TBX21 (D6N8B) XP® Rabbit mAb	H	13051 H	4486 H
2569	TCF4/TCF7L2 (C48H11) Rabbit mAb	H, (M, C)	5111 H	4486 H
2565	TCF4/TCF7L2 (C9B9) Rabbit mAb	H, (M, R)	5111 H	4486 H
92529	TET2 (D9K3E) Rabbit mAb (Mouse Specific)	M	89052 M	8985 M
37785	TFEB (D2O7D) Rabbit mAb	H	27131 H	4486 H
12265	TH1L (D5G6W) Rabbit mAb	H, M, R, Mk, (Hm, B, Dg, Hr)	12558 H	4486 H
12550	Vitamin D3 Receptor (D2K6W) Rabbit mAb	H, M, (Hm, B, Pg, Hr)	25661 H	4486 H
13105	WDR5 (D9E1I) Rabbit mAb	H, M, R, Mk	4641 H	4486 H
14074	YAP (D8H1X) XP® Rabbit mAb	H, M, R, Hm, Mk, (B, GP, Hr)	14927 H	14928 H
14910	BRD7 (D9K2T) Rabbit mAb (ChIP Formulated)	H	12531 H, 9673 H	4486 H
58613	FoxA1/HNF3α (D7P9B) Rabbit mAb	H, M, R, Mk	32784 H	4486 H
83700	NKX3.1 (D2Y1A) XP® Rabbit mAb	H	84846 H	4486 H
62370	SMRT (D8D2L) Rabbit mAb	H, Mk	25661 H	4486 H
18950	TET2 (D6B9Y) Rabbit mAb	H	25946 H	4486 H
订阅新闻

这些引物组是我们内部用于验证ChIP抗体的。阳性对照引物组扩增已显示含有指定组蛋白修饰或被指定转录因子结合的基因位置，而阴性对照引物组不扩增含指定修饰或结合因子的区域。请注意，在指定基因位点发现的组蛋白修饰和转录因子可以根据细胞背景而不同，因此一些对照引物组可能对指定的细胞类型和/或处理具有特异性，这取决于所研究的组蛋白修饰或转录因子。然而，其他对照引物组更通用，并且可以用作多种细胞类型中的多种抗体的对照。例如，RPL30和GAPDH是在大多数细胞类型中有活性的持家基因，因此，RPL30和GAPDH对照引物组可用作在大多数细胞类型中与转录激活相关的组蛋白修饰的阳性对照。类似地，α卫星重复对照引物＃4486 扩增组成型异染色质的区域，并且可以用作组蛋白修饰与任何细胞类型中的转录激活相关的转录因子的阴性对照。

  

【参考文献】

Yulia Haim, Tanya Tarnovscki,Dana Bashari, and Assaf Rudich. A chromatin immunoprecipitation (ChIP) protocol for use in whole human adipose tissue. Am J Physiol Endocrinol Metab 305: E1172–E1177, 2013.

【致谢】

特别感谢Cell Signaling Technology 美国研发科学家陈芳提供的实验数据！

-The End-

首图来自网络