| 货号 | 11966T |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC/ChIP |
| 目标/特异性 | BRM (D9E8B) XP® Rabbit mAb recognizes endogenous levels of total BRM protein. This antibody does not cross-react with BRG1 protein. |
| 使用方法 | WB(1:1000) IP (1:50) IF-IC (1:2400) ChIP (1:100) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | ATP-dependent chromatin remodeling complexes play an essential role in the regulation of various nuclear processes, such as gene expression, DNA replication, and repair (1,2). The SWI/SNF chromatin remodeling complex consists of more than 10 subunits with a single molecule of the ATPase catalytic subunit BRM or BRG1, but not both. The activities of these two subunits drive the disruption of histone-DNA contacts that lead to changes in accessibility of crucial regulatory elements within chromatin (2-5). The BRM/BRG1 containing SWI/SNF complexes are recruited to target promoters by transcription factors, such as nuclear receptors, p53, RB, and BRCA1 to regulate gene activation, cell growth, the cell cycle, and differentiation processes (1,6-9). BRM and BRG1 are also considered to be tumor suppressors and their expression levels are severely reduced in several cancer cell lines (10-13). |
| 存放说明 | -20C |
| 计算分子量 | 200 |
| 参考文献 | 1 . Ho, L. and Crabtree, G.R. (2010) Nature 463, 474-84. 2 . Becker, P.B. and Hörz, W. (2002) Annu Rev Biochem 71, 247-73. 3 . Eberharter, A. and Becker, P.B. (2004) J Cell Sci 117, 3707-11. 4 . Bowman, G.D. (2010) Curr Opin Struct Biol 20, 73-81. 5 . Gangaraju, V.K. and Bartholomew, B. (2007) Mutat Res 618, 3-17. 6 . Lessard, J.A. and Crabtree, G.R. (2010) Annu Rev Cell Dev Biol 26, 503-32. 7 . Morettini, S. et al. (2008) Front Biosci 13, 5522-32. 8 . Wolf, I.M. et al. (2008) J Cell Biochem 104, 1580-6. 9 . Simone, C. (2006) J Cell Physiol 207, 309-14. 10 . Yamamichi, N. et al. (2005) Oncogene 24, 5471-81. 11 . Reisman, D.N. et al. (2002) Oncogene 21, 1196-207. 12 . Shen, H. et al. (2008) Cancer Res 68, 10154-62. 13 . Weissman, B. and Knudsen, K.E. (2009) Cancer Res 69, 8223-30. |
Confocal immunofluorescent analysis of HeLa (positive, left) and NCCIT (negative, right) cells using BRM (D9E8B) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). 采用共聚焦免疫荧光术检测Hela细胞（阳性，左）、NCCIT细胞（阳性，中）以及SirT1 KO MEF 细胞(右) ，使用的抗体为BRM (D9E8B) XP® Rabbit mAb 兔单抗(绿色)。β-Actin (8H10D10) Mouse mAb #3700鼠单抗进行标记（红色）。 | |
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 followed by treatment with β-estradiol (10 nM, 45 min) and either 5 μl of BRM (D9E8B) XP® Rabbit mAb 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 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. 染色质免疫沉淀反应利用4×106MCF细胞进行染色质交联。MCF细胞在含有5%活性炭去除FBS的无酚红培养基中生长4天后，利用β-雌二醇(10nm, 45min) 处理，同时利用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003试剂盒进行分析，采用抗体或为5ul BRM (D9E8B) XP® Rabbit mAb 或为2ul Normal Rabbit IgG #2729。利用SimpleChIP® 人ESR1启动子引物#9673, SimpleChIP® 人PS2启动子引物 #9702, and SimpleChIP® 人α Satellite repeat 引物#4486对DNA含量进行实时PCR.每个样品中免疫沉淀的DNA的量以相对于总的染色质量的比值呈现。 | |
Western blot analysis of extracts from various cell lines using BRM (D9E8B) XP® Rabbit mAb (upper) or Brg1 (A52) Antibody #3508 (lower). Western blot检测各种细胞系提取物,使用抗体BRM (D9E8B) XP® Rabbit mAb 兔单抗(上) or Brg1 (A52) Antibody #3508 (下). |
