Revision 7

#12354Store 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, IHC-P, ChIP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

270

Source/Isotype:

Rabbit IgG

UniProt ID:

#O14497

Entrez-Gene Id:

8289

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
Immunohistochemistry (Paraffin) 1:500 - 1:2000
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

ARID1A/BAF250A (D2A8U) Rabbit mAb recognizes endogenous levels of total ARID1A/BAF250A protein. This antibody also cross-reacts with proteins of unknown origin at 65 kDa.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly1293 of human ARID1A/BAF250A 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).
ARID1A is one of the accessory subunits of the SWI/SNF complex that is an essential part of the esBAF (mouse embryonic stem cell specific SWI/SNF complex). ARID1A is critical for ES cell pluripotency and differentiation into mesoderm-derived cardiomyocytes and adipocytes (10). In addition, ARID1A has been found to be frequently mutated in several cancers such as uterine and ovarian endometrioid carcinoma and ovarian clear cell carcinoma (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. Guan, B. et al. (2011) Am J Surg Pathol 35, 625-32.
  11. Gao, X. et al. (2008) Proc Natl Acad Sci U S A 105, 6656-61.
  12. Jones, S. et al. (2010) Science 330, 228-31.
  13. Wiegand, K.C. et al. (2010) N Engl J Med 363, 1532-43.

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 IHC-P: Immunohistochemistry (Paraffin) 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.
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 7
#12354

ARID1A/BAF250A (D2A8U) Rabbit mAb

Western Blotting Image 1: ARID1A/BAF250A (D2A8U) Rabbit mAb Expand Image
使用 ARID1A/BAF250A (D2A8U) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
Western Blotting Image 2: ARID1A/BAF250A (D2A8U) Rabbit mAb Expand Image
使用 ARID1A/BAF250A (D2A8U) Rabbit mAb(上)或 ß-Actin (D6A8) Rabbit mAb #8457(下)对 T-47D 和 Jurkat 细胞提取物进行蛋白质印迹分析。135kDa-250kDa 之间的某些细胞提取物中可能检测到其他 ARID1A/BAF250A 降解产物,而在 ARID1A/BAF250A 阴性 T-47D 细胞系中缺失这些降解产物。
Immunohistochemistry Image 1: ARID1A/BAF250A (D2A8U) Rabbit mAb Expand Image
使用 ARID1A/BAF250A (D2A8U) Rabbit mAb 对小鼠结肠进行免疫组织化学分析。
Immunohistochemistry Image 2: ARID1A/BAF250A (D2A8U) Rabbit mAb Expand Image
使用 ARID1A/BAF250A (D2A8U) Rabbit mAb 对石蜡包埋的细胞沉淀物、COS-7(左)或 T-47D(右)进行免疫组织化学分析。
Immunohistochemistry Image 3: ARID1A/BAF250A (D2A8U) Rabbit mAb Expand Image
在对照肽(左)或抗原特异性肽(右)存在的情况下,使用 ARID1A/BAF250A (D2A8U) Rabbit mAb 对石蜡包埋的人肺腺鳞癌进行免疫组织化学分析。
Chromatin Immunoprecipitation Image 1: ARID1A/BAF250A (D2A8U) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 NCCIT 细胞的交联染色质和 ARID1A/BAF250A (D2A8U) Rabbit mAb 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。图片中显示 Oct-4/POU5F1 内结合,而 Oct-4/POU5F1 是已知的 ARID1A 靶标基因(请参阅含有 ChIP-qPCR 数据的附图)。
Chromatin Immunoprecipitation Image 2: ARID1A/BAF250A (D2A8U) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 NCCIT 细胞的交联染色质和 ARID1A/BAF250A (D2A8U) Rabbit mAb 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。该图显示在染色体 6(上图)内的结合,包括 ARID1A 的已知靶标基因 Oct-4/POU5F1(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: ARID1A/BAF250A (D2A8U) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005,对 NCCIT 细胞中提取的交联染色质,在加入 ARID1A/BAF250A (D2A8U) Rabbit mAb 或 Normal Rabbit IgG #2729 后进行染色质免疫沉淀。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。