Revision 5

#23064Store 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, IF-F, IF-IC, FC-FP, ChIP, ChIP-seq, C&R

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

35

Source/Isotype:

Rabbit IgG

UniProt ID:

#P48431

Entrez-Gene Id:

6657

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 1:100
Immunofluorescence (Frozen) 1:400
Immunofluorescence (Immunocytochemistry) 1:400
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800
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

Sox2 (D9B8N) Rabbit mAb recognizes endogenous levels of total Sox2 protein. The abundant nonspecific cytoplasmic labeling was observed in adult brain by immunofluorescence (IF). However, the specific staining was observed in embryonic tissue, including brain, by IF.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala188 of human Sox2 protein.

Background

Embryonic stem cells (ESC) derived from the inner cell mass of the blastocyst are unique in their pluripotent capacity and potential for self-renewal (1). Research studies demonstrate that a set of transcription factors that includes Oct-4, Sox2, and Nanog forms a transcriptional network that maintains cells in a pluripotent state (2,3). Chromatin immunoprecipitation experiments show that Sox2 and Oct-4 bind to thousands of gene regulatory sites, many of which regulate cell pluripotency and early embryonic development (4,5). siRNA knockdown of either Sox2 or Oct-4 results in loss of pluripotency (6). Induced overexpression of Oct-4 and Sox2, along with additional transcription factors Klf4 and c-Myc, can reprogram both mouse and human somatic cells to a pluripotent state (7,8). Additional evidence demonstrates that Sox2 is also present in adult multipotent progenitors that give rise to some adult epithelial tissues, including several glands, the glandular stomach, testes, and cervix. Sox2 is thought to regulate target gene expression important for survival and regeneration of these tissues (9).

  1. Conley, B.J. et al. (2004) Int J Biochem Cell Biol 36, 555-67.
  2. Pesce, M. and Schöler, H.R. (2001) Stem Cells 19, 271-8.
  3. Pan, G. and Thomson, J.A. (2007) Cell Res 17, 42-9.
  4. Boyer, L.A. et al. (2005) Cell 122, 947-56.
  5. Loh, Y.H. et al. (2006) Nat Genet 38, 431-40.
  6. Matin, M.M. et al. (2004) Stem Cells 22, 659-68.
  7. Takahashi, K. and Yamanaka, S. (2006) Cell 126, 663-76.
  8. Okita, K. et al. (2007) Nature 448, 313-7.
  9. Arnold, K. et al. (2011) Cell Stem Cell 9, 317-29.

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 IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) 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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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
#23064

Sox2 (D9B8N) Rabbit mAb

Western Blotting Image 1: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 Sox2 (D9B8N) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: Sox2 (D9B8N) Rabbit mAb Expand Image
对F9 细胞提取物 Sox2 进行免疫沉淀。泳道 1 为10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Sox2 (D9B8N) Rabbit mAb。使用 Sox2 (D9B8N) Rabbit mAb 进行蛋白质印迹分析。
Immunofluorescence Image 1: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 Sox2 (D9B8N) Rabbit mAb (绿色)对发育中的小鼠 E12 前肠(左图)、脊髓和体节(中图)或脑(右图)进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Immunofluorescence Image 1: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 Sox2 (D9B8N) Rabbit mAb (绿色)对 F9(左图)、3T3(中左图)、NCCIT(中右图) 或 HeLa(右图)细胞进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。
Flow Cytometry Image 1: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 Sox2 (D9B8N) Rabbit mAb 对 Hela 细胞(蓝色)和 NTERA 细胞(绿色)进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
Flow Cytometry Image 2: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 Sox2 (D9B8N) Rabbit mAb,对 NIH3T3 细胞(蓝色)和 F9 细胞(绿色)进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
Chromatin Immunoprecipitation Image 1: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 mES 细胞的交联染色质和 Sox2 (D9B8N) 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。结果图显示在 POU5F1/OCT4 内结合,POU5F1/OCT4 是一种已知的 Sox2 靶标基因(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 2: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 mES 细胞的交联染色质和 Sox2 (D9B8N) 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。结果图显示在染色体 17(上图)内的结合,包括 Sox2 的已知靶基因 POU5F1/OCT4(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
Chromatin Immunoprecipitation Image 3: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,对 mES 细胞中提取的交联染色质,在加入 Sox2 (D9B8N) Rabbit mAb 或 Normal Rabbit IgG #​2729 后,进行染色质免疫沉淀。使用 SimpleChIP® Mouse Oct-4 Promoter Primers #4653、SimpleChIP® Mouse XIST Intron 1 Primers #4659 和 SimpleChIP® Mouse RPL30 Intron 2 Primers #7015,通过实时 PCR 的方法对富集的 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: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,NCCIT 细胞和 Sox2 (D9B8N) 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 Nanog 内结合,Nanog 是一种已知的 Sox2 靶标基因(参见包含 CUT&RUN-qPCR 数据的其他图形)。
CUT and RUN Image 2: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,NCCIT 细胞和 Sox2 (D9B8N) 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 12(上图)内的结合,包括 Sox2 的已知靶标基因 Nanog(下图)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: Sox2 (D9B8N) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,NCCIT 细胞和 Sox2 (D9B8N) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362进行 CUT&RUN。使用 SimpleChIP® Human Nanog Promoter Primers #95064、SimpleChIP® Human Oct-4 Promoter Primers #4641 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。