Revision 1

#89123Store 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, IP, ChIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

205

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q86U86

Entrez-Gene Id:

55193

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 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
Immunoprecipitation 1:50
Chromatin IP 1:50

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

PBRM1/BAF180 (E9X2Z) Rabbit mAb recognizes endogenous levels of total PBRM1/BAF180 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu1672 of human PBRM1/BAF180 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).

PBRM1/BAF180 is a unique member of the SWI/SNF complex PBAF, which binds to kinetochores in mitotic chromatin (10,11). PBAF is involved in nuclear receptor-mediated transcription and retinoic acid driven gene activation (12,13). PBRM1/BAF180 has been shown to be a potent tumor suppressor, as it is the second-most mutated gene in renal carcinomas (14). Mutations of PBRM1/BAF180 have also been shown to be involved in breast cancer, and low expression relates to poorer prognosis (15,16). PBRM1/BAF180 is phosphorylated at Ser948 by ATM during DNA damage, which is important for transcriptional silencing and repair around double-stranded breaks (17).

  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. Nie, Z. et al. (2000) Mol Cell Biol 20, 8879-88.
  11. Xue, Y. et al. (2000) Proc Natl Acad Sci U S A 97, 13015-20.
  12. Lemon, B. et al. (2001) Nature 414, 924-8.
  13. Wang, Z. et al. (2004) Genes Dev 18, 3106-16.
  14. Varela, I. et al. (2011) Nature 469, 539-42.
  15. Xia, W. et al. (2008) Cancer Res 68, 1667-74.
  16. Mo, D. et al. (2015) Int J Clin Exp Pathol 8, 9307-13.

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 IP: Immunoprecipitation 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.
CST is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered 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 1
#89123

PBRM1/BAF180 (E9X2Z) Rabbit mAb

Western Blotting Image 1: PBRM1/BAF180 (E9X2Z) Rabbit mAb Expand Image
使用 PBRM1/BAF180 (E9X2Z) Rabbit mAb(上图)或 α-Actinin (D6F6) XP® Rabbit mAb #6487(下图)对不同细胞系的提取物进行蛋白印迹分析。正如预期的一样,HCC1143 细胞的 PBRM1/BAF180 表达呈阴性。
Immunoprecipitation Image 1: PBRM1/BAF180 (E9X2Z) Rabbit mAb Expand Image
对 Jurkat 细胞提取物的 PBRM1/BAF180 蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 PBRM1/BAF180 (E9X2Z) Rabbit mAb。使用 PBRM1/BAF180 (E9X2Z) Rabbit mAb 进行蛋白印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Chromatin Immunoprecipitation Image 1: PBRM1/BAF180 (E9X2Z) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对经无酚红培养基和 5% 炭剥离的 FBS 中培养 4 天,随后用 β-雌二醇(100 nM)持续处理 45 分钟的 MCF7 细胞中提取的交联染色质,在加入 PBRM1/BAF180 (E9X2Z) Rabbit mAb 或 Normal Rabbit IgG #2729 ​后,进行染色质免疫沉淀分析。使用 Human ESR1 Intron 1 Primers、SimpleChIP® Human CCND1 Promoter Primers #12531 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。