Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

PRMT4/CARM1 (3H2) Mouse mAb #12495

No. Size Price
12495S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
12495 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 63 Mouse IgG1
IP 1:100
IF-IC 1:100
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

PRMT4/CARM1 (3H2) Mouse mAb recognizes endogenous levels of total PRMT4/CARM1 protein.

PRMT4/CARM1 (3H2) Mouse mAb鼠单抗能够检测内源性PRMT4/CARM1 总蛋白水平。

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the human CARM1 protein.

该单克隆抗体经是采用合成的人CARM1重组蛋白免疫动物而生产的。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using PRMT4/CARM1 (3H2) Mouse mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).采用共聚焦免疫荧光术检测HeLa细胞,使用的抗体为PRMT4/CARM1 (3H2) Mouse mAb (绿色)。肌动蛋白微丝使用Alexa Fluor® 488 Phalloidin #8878进行标记(红色)。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using PRMT4/CARM1 (3H2) Mouse mAb.Western blot方法检测不同细胞系的提取物,使用的抗体为 PRMT4/CARM1 (3H2) Mouse mAb。

IP

IP

Immunoprecipitation of PRMT4/CARM1 from HCT 116 cell extracts using Mouse (G3A1) mAb IgG1 Isotype control #5415 (lane 2) or PRMT4/CARM1 (3H2) Mouse mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using PRMT4/CARM1 (C31G9) Rabbit mAb #3379.从 HCT 116细胞提取物中免疫沉淀 PRMT4/CARM1,使用的抗体为Mouse (G3A1) mAb IgG1 Isotype control #5415 (lane 2)或 PRMT4/CARM1 (3H2) Mouse mAb (lane 3)。Lane 1为10% input。 使用PRMT4/CARM1 (C31G9) Rabbit mAb #3379进行Western blot检测。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 minutes and either 10 μl of PRMT4/CARM1 (3H2) or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CCND1 Promoter Primers #12531, SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.使用从4 x 106 MCF7细胞中提取的交联过的染色质,以及10 µl PRMT4/CARM1 (3H2) 或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀,这些细胞需要在含5%活性炭处理胎牛血清以及无酚红的培养基中培养4天,之后再用β-estradiol 进行处理(10 nM, 45 min)。所用试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。富集到的DNA经过实时PCR定量,所使用引物为SimpleChIP® Human CCND1 Promoter Primers #12531, SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702,以及SimpleChIP® Human α Satellite Repeat Primers #4486。每个样品中免疫沉淀得到的DNA量由与input chromatin( 相当于1)的相对信号值来表示。

Background

Protein arginine N-methyltransferase 1 (PRMT1) is a member of the protein arginine N-methyltransferase (PRMT) family of proteins that catalyze the transfer of a methyl group from S-adenosylmethionine (AdoMet) to a guanidine nitrogen of arginine (1). Though all PRMT proteins catalyze the formation of mono-methyl arginine, Type I PRMTs (PRMT1, 3, 4, and 6) add an additional methyl group to produce an asymmetric di-methyl arginine while Type II PRMTs (PRMT 5 and 7) produce symmetric di-methyl arginine (1). Mono-methyl arginine, but not di-methyl arginine, can be converted to citrulline through deimination catalyzed by enzymes such as PADI4 (2). Most PRMTs, including PRMT1, methylate arginine residues found within glycine-arginine rich (GAR) protein domains, such as RGG, RG, and RXR repeats (1). However, PRMT4/CARM1 and PRMT5 methylate arginine residues within PGM (proline-, glycine-, methionine-rich) motifs (3). PRMT1 methylates Arg3 of histone H4 and cooperates synergistically with p300/CBP to enhance transcriptional activation by nuclear receptor proteins (4-6). In addition, PRMT1 methylates many non-histone proteins, including the orphan nuclear receptor HNF4 (6), components of the heterogeneous nuclear ribonucleoprotein (hnRNP) particle (7), the RNA binding protein Sam68 (8), interleukin enhancer-binding factor 3 (ILF3) (9) and interferon-α and β receptors (10). These interactions suggest additional functions in transcriptional regulation, mRNA processing and signal transduction. Alternative mRNA splicing produces three enzymatically active PMRT1 isoforms that differ in their amino-terminal regions (11). PRMT1 is localized to the nucleus or cytoplasm, depending on cell type (12,13), and appears in many distinct protein complexes. ILF3, TIS21 and the leukemia-associated BTG1 proteins bind PRMT1 to regulate its methyltransferase activity (9,14).

Protein arginine N-methyltransferase 4 (PRMT4)是蛋白精氨酸甲基转移酶(PRMT)蛋白家族的一个成员,它以S-腺苷-甲硫氨酸(S-adenosylmethionine,AdoMet)为甲基供体,把甲基转移到蛋白精氨酸胍基的氮原子上 (1)。尽管所有的PRMT蛋白都能催化单甲基精氨酸的形成,但Type I PRMTs (PRMT1, 3, 4, 和 6)能够添加另一个甲基基团从而产生一个不对称的二甲基化精氨酸,而Type II PRMTs (PRMT 5 and 7)则能产生一个对称的二甲基精氨酸(1)。单甲基化精氨酸,而非二甲基化精氨酸可以通过由类似PADI4这样的酶催化的脱氨基作用来转变为瓜氨酸(2)。大部分PRMT包括PRMT1,能够甲基化在富含甘氨酸 - 精氨酸(GAR)的蛋白结构域内发现的精氨酸残基,例如RGG, RG, 和 RXR重复序列中 (1)。然而,PRMT4/CARM1 和 PRMT5可以使PGM (proline-, glycine-, methionine-rich) 结构中精氨酸甲基化(3)。PRMT1能够甲基化组蛋白H4的Arg3 位点,并且能够与p300/CBP协同作用通过核受体蛋白来增强转录激活(4-6)。此外,PRMT1能够甲基化组非组蛋白,包括孤儿核受体HNF4(6)、不均一核糖核蛋白(hnRNP)颗粒的组分(7)、RNA结合蛋白Sam68(8)、白介素增强子结合因子3(ILF3)(9)以及干扰素-α 和 β受体(10)。这些相互作用暗示着PRMT1在转录调控、mRNA加工和信号转导还有一些其他的功能。可变RNA剪接会产生3种氨基末端区域不一样的酶促激活PMRT1异构体(11)。PRMT1根据不同的细胞类型会定位在和或者细胞质中(12,13),同时会在许多不同的蛋白复合物中出现。ILF3、TIS21以及白血病相关性BTG1蛋白能够结合PRMT1从而条龙其甲基转移酶活性(9,14)。

  1. Bedford, M.T. and Richard, S. (2005) Mol. Cell 18, 263-272.
  2. Wang, Y. et al. (2004) Science 306, 279-283.
  3. Cheng, D. et al. (2007) Mol. Cell 25, 71-83.
  4. Wang, H. et al. (2001) Science 293, 853-857.
  5. Strahl, B.D. et al. (2001) Curr. Biol. 11, 996-1000.
  6. Barrero, M.J. and Malik, S. (2006) Mol. Cell 24, 233-243.
  7. Nichols, R.C. et al. (2000) Exp. Cell Res. 256, 522-532.
  8. Côté, J. et al. (2003) Mol. Biol. Cell 14, 274-287.
  9. Tang, J. et al. (2000) J. Biol. Chem. 275, 19866-19876.
  10. Abramovich, C. et al. (1997) EMBO J. 16, 260-266.
  11. Scorilas, A. et al. (2000) Biochem. Biophys. Res. Commun. 278, 349-359.
  12. Frankel, A. et al. (2002) J. Biol. Chem. 277, 3537-3543.
  13. Herrmann, F. et al. (2005) J. Biol. Chem. 280, 38005-38010.
  14. Lin, W.J. et al. (1996) J. Biol. Chem. 271, 15034-15044.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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