Revision 5

#91735Store 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, IHC-P, ChIP, ChIP-seq, C&R

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

44

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q12824

Entrez-Gene Id:

6598

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.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
Immunohistochemistry (Paraffin) 1:500 - 1:2000
Chromatin IP 1:50
Chromatin IP-seq 1:50
CUT&RUN 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

SMARCB1/BAF47 (D8M1X) Rabbit mAb recognizes endogenous levels of total SMARCB1/BAF47 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu120 of human SMARCB1/BAF47 protein.

Background

ATP-dependent chromatin remodeling complexes play an essential role in the regulation of nuclear processes such as transcription and DNA replication and repair (1,2). The SWI/SNF chromatin remodeling complex consists of more than 10 subunits and contains a single molecule of either BRM or BRG1 as the ATPase catalytic subunit. The activity of the ATPase subunit disrupts histone-DNA contacts and changes the accessibility of crucial regulatory elements to the chromatin. The additional core and accessory subunits play a scaffolding role to maintain stability and provide surfaces for interaction with various transcription factors and chromatin (2-5). The interactions between SWI/SNF subunits and transcription factors, such as nuclear receptors, p53, Rb, BRCA1, and MyoD, facilitate recruitment of the complex to target genes for regulation of gene activation, cell growth, cell cycle, and differentiation processes (1,6-9).
SMARCB1/BAF47, one of the core subunits of the SWI/SNF complex, is necessary for efficient nucleosome remodeling by BRG1 in vitro (10). SMARCB1/BAF47 is an essential part of the esBAF (mouse embryonic stem cell specific SWI/SNF complex) and is necessary for early embryogenesis and hepatocyte differentiation (11,12). In addition, SMARCB1/BAF47 is considered to be a tumor suppressor protein; inactivating mutations have been indentified in a large number of malignant rhabdoid tumors (13,14).

  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. Phelan, M.L. et al. (1999) Mol Cell 3, 247-53.
  11. Klochendler-Yeivin, A. et al. (2000) EMBO Rep 1, 500-6.
  12. Gresh, L. et al. (2005) EMBO J 24, 3313-24.
  13. Versteege, I. et al. (1998) Nature 394, 203-6.
  14. Biegel, J.A. et al. (1999) Cancer Res 59, 74-9.

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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN

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.
SignalStain 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 5
#91735

SMARCB1/BAF47 (D8M1X) Rabbit mAb

Western Blotting Image 1: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
使用 SMARCB1/BAF47 (D8M1X) Rabbit mab(上图)或 β-Actin (13E5) Rabbit mAb #4970(下图),对不同细胞系的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
对 293T 细胞提取物 SMARCB1/BAF47 进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb XP® Isotype Control #3900,泳道 3 为 SMARCB1/BAF47 (D8M1X) Rabbit mAb。使用 SMARCB1/BAF47 (D8M1X) Rabbit mAb 进行蛋白质印迹分析。
Immunohistochemistry Image 1: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
使用 SMARCB1/BAF47 (D8M1X) Rabbit mAb 对石蜡包埋的人结肠癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
使用 SMARCB1/BAF47 (D8M1X) Rabbit mAb 对石蜡包埋的小鼠结肠细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
使用 SMARCB1/BAF47 (D8M1X) Rabbit mAb 对石蜡包埋的子宫内膜腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
使用 SMARCB1/BAF47 (D8M1X) Rabbit mAb 对石蜡包埋的 HeLa (左图)和 A204 (右图)细胞沉淀物进行免疫组织化学分析。
Immunohistochemistry Image 5: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
在有对照肽(左图)或抗原特异性肽(右图)的情况下,使用 SMARCB1/BAF47 (D8M1X) Rabbit mAb 对石蜡包埋的人肺癌细胞进行免疫组织化学分析。
Chromatin Immunoprecipitation Image 1: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
使用 SimpleChIP® Plus 酶解试剂盒 (磁珠法) #9005 对在无酚红的培养基和 5% 活性炭剥离的 FBS 中生长 4 天,随后使用 β-雌二醇(10 nM,45 分钟)处理的 MCF7 细胞的交联染色质与 SMARCC1/BAF155 (D7F8S) 兔单克隆抗体 #11956、SMARCB1/BAF47 (D8M1X) 兔单克隆抗体 #91735 或 SS18 (D6I4Z) 兔单克隆抗体 #21792 进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。SMARCC1/BAF155、SMARCB1/BAF47 和 SS18 均为 SWI/SNF 复合体的所有亚基。结果图显示了 pS2/TFF1 内结合,pS2/TFF1 是一种已知的 SWI/SNF 靶标基因(参见包含 ChIP-qPCR 数据的其他结果图)。如需了解其他 ChIP-seq 情况,请下载产品说明书。
Chromatin Immunoprecipitation Image 2: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,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 SMARCC1/BAF155 (D7F8S) Rabbit mAb #11956 或 SS18 (D6I4Z) Rabbit mAb #21792 后,进行染色质免疫沉淀。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。SMARCC1/BAF155、SMARCB1/BAF47 和 SS18 均为 SWI/SNF 复合体的所有亚基。结果图显示在染色体21(上图)内的结合,包括 SWI/SNF 复合体的已知靶标基因 pS2/TFF1(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: SMARCB1/BAF47 (D8M1X) 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 SMARCB1/BAF47 (D8M1X) Rabbit mAb 或 Normal Rabbit IgG #2729 后进行染色质免疫沉淀。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。
CUT and RUN Image 1: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
对在无酚红培养基和 5% 活性炭剥离 FBS 中生长 4 天后接着使用 β 雌二醇(10 nM,45 分钟)进行处理的 MCF7 细胞和 SMARCB1/BAF47 (D8M1X) Rabbit mAb,通过 CUT&RUN Assay Kit #86652 进行 CUT&RUN 分析。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该结果图显示在已知 SMARCB1 靶标基因 TFF1 内的结合(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 2: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
对在无酚红培养基和 5% 活性炭剥离 FBS 中生长 4 天后接着使用 β 雌二醇(10 nM,45 分钟)进行处理的 MCF7 细胞和 SMARCB1/BAF47 (D8M1X) Rabbit mAb,通过 CUT&RUN Assay Kit #86652 进行 CUT&RUN 分析。使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。结果图显示在染色体 21(上图)内的结合,包括 SMARCB1 的已知靶标基因 TFF1(下图)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: SMARCB1/BAF47 (D8M1X) Rabbit mAb Expand Image
对在无酚红培养基和 5% 活性炭剥离 FBS 中生长 4 天后接着使用 β 雌二醇(10 nM,45 分钟)进行处理的 MCF7 细胞和 SMARCB1/BAF47 (D8M1X) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362,通过 CUT&RUN Assay Kit #86652 进行 CUT&RUN 分析。使用 SimpleChIP® Human ESR1 Promoter Primers #9673、SimpleChIP® Human pS2 Promoter Primers #9702 和 SimpleChIP® Human α Satellite Repeat Primers #4486 进行实时 PCR 来对富集的 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。