Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

BRM (D9E8B) XP® Rabbit mAb #11966

sc-28710   sc-6450   SWI/SNF  

No. Size Price
11966S 100 µl ( 10 western blots ) ¥3,580.00 现货查询 购买询价
11966T 20 µl ( 2 western blots ) ¥1,400.00 现货查询 购买询价
11966 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Monkey, Endogenous 200 Rabbit IgG
IP 1:50
IF-IC 1:2400
ChIP 1:100

Species cross-reactivity is determined by western blot.

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

Homology

Species predicted to react based on 100% sequence homology: Dog,

Specificity / Sensitivity

BRM (D9E8B) XP® Rabbit mAb recognizes endogenous levels of total BRM protein. This antibody does not cross-react with BRG1 protein.

BRM (D9E8B) XP® Rabbit mAb兔单抗识别内源性BRM总蛋白水平。该抗体不与BRG1蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly264 of human BRM protein.

该该单克隆抗体是采用合成的与人源BRM蛋白Gly264残基周围序列相对应的肽段免疫动物而生产的。

IF-IC

IF-IC

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 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 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 Blotting

Western Blotting

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).

Background

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).

ATP依赖的染色质重塑复合物,在调节如转录、DNA的复制和修复等细胞核反应起着重要作用(1,2)。SWI/SNF染色质重塑复合物由10多个亚基组成,BRM或BRG1任一单分子作为ATP酶催化亚基,但是两者不能同时存在。BRM/BRG1能够阻断组蛋白与DNA的结合,改变染色质内重要的辅助调控元件(2-5)。含有BRM/BRG1的SWI/SNF复合物招募一些转录因子靶向启动子,如细胞核受体、P53、RB和BRCA1,从而调节细胞生长、细胞周期、细胞分化等过程(1,6-9)。BRM及BRG1也被认为是肿瘤抑制基因,在几种癌症细胞中,两者的表达水平严重降低(10-13)。

  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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products


For Research Use Only. Not For Use In Diagnostic Procedures.

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

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

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

用户评论 --- 共 0

该产品暂无评论!

我要参与评论 :

如要参与评论请先登录网站

还没有网站账户?去注册一下吧

Products

 

Applications