Revision 9

#11956Store 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, ChIP-seq, C&R, C&T

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

155

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q92922

Entrez-Gene Id:

6599

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq 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.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

The CUT&Tag dilution was determined using CUT&Tag Assay Kit #77552.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Chromatin IP 1:100
Chromatin IP-seq 1:100
CUT&RUN 1:100
CUT&Tag 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

SMARCC1/BAF155 (D7F8S) Rabbit mAb recognizes endogenous levels of total SMARCC1/BAF155 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly975 of human SMARCC1/BAF155 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).

SMARCC1/BAF155 is one of the core subunits of the SWI/SNF complex, which is necessary for efficient nucleosome remodeling by BRG1 in vitro (10). SMARCC1 is an essential part of the mouse embryonic stem cell specific SWI/SNF complex (esBAF), which is necessary for early embryogenesis, especially proper brain and visceral endoderm development (11-13).

  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. Phelan, M.L. et al. (1999) Mol Cell 3, 247-53.
  11. Han, D. et al. (2008) Dev Biol 315, 136-46.
  12. Kim, J.K. et al. (2001) Mol Cell Biol 21, 7787-95.
  13. Schaniel, C. et al. (2009) Stem Cells 27, 2979-91.

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 ChIP-seq: Chromatin IP-seq C&R: CUT&RUN C&T: CUT&Tag

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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。
Revision 9
#11956

SMARCC1/BAF155 (D7F8S) Rabbit mAb

Western Blotting Image 1: SMARCC1/BAF155 (D7F8S) Rabbit mAb Expand Image
使用 SMARCC1/BAF155 (D7F8S) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: SMARCC1/BAF155 (D7F8S) Rabbit mAb Expand Image
使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900（泳道 2）或 SMARCC1/BAF155 (D7F8S) Rabbit mAb （泳道 3），对 HeLa 细胞提取物 SMARCC1/BAF155 进行免疫沉淀。泳道 1 是 10% 输入对照。使用 SMARCC1/BAF155 (D7F8S) Rabbit mAb 进行蛋白质印迹分析。
Chromatin Immunoprecipitation Image 1: SMARCC1/BAF155 (D7F8S) 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，在加入 SMARCC1/BAF155 (D7F8S) Rabbit mAb, SMARCB1/BAF47 (D8M1X) Rabbit mAb #91735 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）。
Chromatin Immunoprecipitation Image 2: SMARCC1/BAF155 (D7F8S) 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，在加入 SMARCC1/BAF155 (D7F8S) Rabbit mAb, SMARCB1/BAF47 (D8M1X) Rabbit mAb #91735 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: SMARCC1/BAF155 (D7F8S) 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，在加入 SMARCC1/BAF155 (D7F8S) 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: SMARCC1/BAF155 (D7F8S) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652，通过 NCCIT 细胞和 SMARCC1/BAF155 (D7F8S) Rabbit mAb 进行 CUT&RUN。‚使用 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。图形显示在 Sox2 内结合，Sox2 是一种已知的 SMARCC1 靶标基因（参见包含 CUT&RUN-qPCR 数据的其他图形）。
CUT and RUN Image 2: SMARCC1/BAF155 (D7F8S) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652，通过 NCCIT 细胞和 SMARCC1/BAF155 (D7F8S) Rabbit mAb 进行 CUT&RUN。‚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 3（上图）内的结合，包括 SMARCC1 的已知靶标基因 Sox2（下图）（参见包含 CUT&RUN-qPCR 数据的其他结果图）。
CUT and RUN Image 3: SMARCC1/BAF155 (D7F8S) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652，NCCIT 细胞和 SMARCC1/BAF155 (D7F8S) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362进行 CUT&RUN。使用 SimpleChIP® Human Oct-4 Promoter Primers #4641、SimpleChIP® Human Sox2 Promoter Primers #4649 和 SimpleChIP® Human α Satellite Repeat Primers #4486，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 Tag Image 1: SMARCC1/BAF155 (D7F8S) Rabbit mAb Expand Image
CUT&Tag was performed with NCCIT cells and SMARCC1/BAF155 (D7F8S) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figure shows binding across SOX2.
CUT and Tag Image 2: SMARCC1/BAF155 (D7F8S) Rabbit mAb Expand Image
CUT&Tag was performed with NCCIT cells and SMARCC1/BAF155 (D7F8S) Rabbit mAb, using CUT&Tag Assay Kit #77552. DNA library was prepared using CUT&Tag Dual Index Primers and PCR Master Mix for Illumina Systems #47415. The figures show binding across chromosome 3 (upper), including SOX2 gene (lower).
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Orders: 877-616-CELL (2355)•[email protected]• Support: 877-678-TECH (8324)•[email protected]• Web: cellsignal.com
仅供研究使用。不得用于诊断流程。