Revision 6

#11966Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, W-S, IP, IHC-P, IF-IC, ChIP

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

200

Source/Isotype:

Rabbit IgG

UniProt ID:

#P51531

Entrez-Gene Id:

6595

Product Information

Product Usage Information

For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:3200 - 1:6400
Chromatin IP 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

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

Species Reactivity:

Human, Monkey

Species predicted to react based on 100% sequence homology

Dog

Source / Purification

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

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).

  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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 6
#11966

BRM (D9E8B) XP® Rabbit mAb

Western Blotting Image 1: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 BRM (D9E8B) XP® Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对对照的 HeLa 细胞(泳道 1)或 BRM 编码基因出现靶向突变的 HeLa 细胞(泳道 2)的提取物进行蛋白质印迹分析。突变型 HeLa 细胞的 BRM 分子量变化证实了抗体对 BRM 的特异性。
Western Blotting Image 2: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 BRM (D9E8B) XP® 兔单抗(上)或 Brg1 (A52) Antibody #3508(下)对来自不同细胞系的提取物进行蛋白质印迹分析。
Western Blotting Image 1: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 BRM (D9E8B) XP® Rabbit mAb #11966 对 A549 细胞的裂解物 (1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
Immunoprecipitation Image 1: BRM (D9E8B) XP® Rabbit mAb Expand Image
对 HeLa 细胞提取物的 BRM 蛋白进行的免疫沉淀分析。泳道 1 为 10 % 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 BRM (D9E8B) XP® Rabbit mAb。使用 BRM (D9E8B) XP® Rabbit mAb 进行蛋白质印迹分析。
Immunohistochemistry Image 1: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 BRM (D9E8B) XP (R) 兔单抗对石蜡包埋的人前列腺进行免疫组织化学分析。
Immunohistochemistry Image 2: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 BRM (D9E8B) XP (R) 兔单抗对石蜡包埋的人结肠癌进行免疫组织化学分析。
Immunohistochemistry Image 3: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 BRM (D9E8B) XP (R) Rabbit mAb 对石蜡包埋的人卵巢透明细胞腺癌进行免疫组织化学分析。
Immunohistochemistry Image 4: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 BRM (D9E8B) XP (R) 兔单抗对石蜡包埋的 HeLa(左)细胞沉淀物和 NCC(右)细胞沉淀物进行免疫组织化学分析。
Immunofluorescence Image 1: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 BRM (D9E8B) XP® Rabbit mAb(绿色)和 β-Actin (8H10D10) Mouse mAb #3700(红色)对 HeLa(阳性,左)细胞和 NCC(阴性,右)细胞进行共聚焦免疫荧光分析。
Chromatin Immunoprecipitation Image 1: BRM (D9E8B) XP® Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对在无酚红的培养基和 5% 活性炭剥离的 FBS 中生长 4 天,然后使用 β-雌二醇(10 nM, 45 分钟)处理的MCF7 细胞中提取的交联染色质,在加入BRM (D9E8B) XP® Rabbit mAb 或 Normal Rabbit IgG #2729后,进行交联染色质免疫沉淀。使用 SimpleChIP® Human ESR1 Promoter Primers #9673、SimpleChIP® Human pS2 Promoter Primers #9702 和 SimpleChIP® Human α Satellite Repeat Primers #4486 并通过实时 PCR 对富集的 DNA 进行定量。将每份样品中免疫沉淀的 DNA 的量表现为相对于所输入染色质总量(等于 1)的信号。