| 货号 | 12495S |
| 反应种属 | Human/Mouse/Rat/Monkey |
| 来源宿主 | Mouse |
| 应用 | W/IP/IF-IC/ChIP |
| 目标/特异性 | PRMT4/CARM1 (3H2) Mouse mAb recognizes endogenous levels of total PRMT4/CARM1 protein. |
| 使用方法 | WB(1:1000) IP (1:100) IF-IC (1:100) ChIP (1:50) |
| 供应商 | CST |
| 背景 | 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). |
| 存放说明 | -20C |
| 计算分子量 | 63 |
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 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。 | |
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 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。 |
