Revision 5

#46395Store 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-IC, FC-FP, ChIP, ChIP-seq, C&R

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

65

Source/Isotype:

Rabbit IgG

UniProt ID:

#P25490

Entrez-Gene Id:

7528

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
Immunofluorescence (Immunocytochemistry) 1:800
Flow Cytometry (Fixed/Permeabilized) 1:100
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

YY1 (D5D9Z) Rabbit mAb recognizes endogenous levels of total YY1 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human YY1 protein. The antibody epitope has been further mapped to a peptide corresponding to residues surrounding Pro284 of human YY1 protein.

Background

YY1 (Yin Yang1) is a ubiquitously expressed transcription factor with fundamental roles in embryogenesis, differentiation, replication and proliferation. YY1 contains four zinc finger motifs of the Cys-Cys-His-His type and can activate different eukaryotic genes (such as CREB, c-myc, Histone H4, p53 and PARP-1) or repress different eukaryotic genes (such as α-actin, IFN-β and IFN-γ) as well as regulate some viral promoters (1). YY1 deficient embryos die approximately at the time of implantation, suggesting that YY1 has an essential role in embryonic development (2). YY1 is overexpressed in cancer cells such as prostate cancer and therefore may be considered a prognostic marker (1).

  1. Gordon, S. et al. (2006) Oncogene 25, 1125-42.
  2. Donohoe, M.E. et al. (1999) Mol. Cell Biol. 19, 7237-7244.

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-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.
The transfer of this product is contingent on the buyer using the purchased product solely in research conducted by the buyer (whether the buyer is an academic or for-profit entity), for Immunocytochemistry, high content screening (HCS) analysis, or flow cytometry applications. The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) resale, whether or not such product or its components are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].
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
#46395

YY1 (D5D9Z) Rabbit mAb

Western Blotting Image 1: YY1 (D5D9Z) Rabbit mAb Expand Image
使用 YY1 (D5D9Z) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: YY1 (D5D9Z) Rabbit mAb Expand Image
对 MOLT-4 细胞提取物 YY1 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 YY1 (D5D9Z) Rabbit mAb。使用 YY1 (D5D9Z) Rabbit mAb 和 Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 进行蛋白质印迹分析。
Immunofluorescence Image 1: YY1 (D5D9Z) Rabbit mAb Expand Image
使用 YY1 (D5D9Z) Rabbit mAb(绿色)对 HeLa 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: YY1 (D5D9Z) Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 YY1 (D5D9Z) Rabbit mAb(实线)对 MCF7 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
Chromatin Immunoprecipitation Image 1: YY1 (D5D9Z) Rabbit mAb Expand Image
使用 SimpleChIP® 酶解染色质免疫共沉淀试剂盒(磁珠法)#9005 对 K-562 细胞的交联染色质与 YY1 (D5D9Z) 兔单克隆抗体或正常兔 IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® CXXC1 Exon 1 Primers #67406、FAM193B promoter primers 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。
Chromatin Immunoprecipitation Image 1: YY1 (D5D9Z) Rabbit mAb Expand Image
使用 SimpleChIP® 酶解染色质免疫共沉淀试剂盒(磁珠法) #9005 对 K-562 细胞的交联染色质和 YY1 (D5D9Z) 兔单克隆抗体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5(上图)内的结合,包括 YY1 的已知靶基因 FAM193B(下图)(参见包含 ChIP-qPCR 数据的其他结果图)。
CUT and RUN Image 1: YY1 (D5D9Z) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以K562 细胞和 YY1 (D5D9Z) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示结合作用遍及 RBM15 基因(YY1 的已知靶标基因) (参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 2: YY1 (D5D9Z) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652,以K562 细胞和 YY1 (D5D9Z) Rabbit mAb 进行 CUT&RUN。使用 DNA Library Prep Kit for Illumina Systems (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示结合作用遍及染色体 #1(上图),包括RBM15(YY1 的已知靶标基因)(下图)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: YY1 (D5D9Z) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 ,以 K562 细胞和 YY1 (D5D9Z) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN。使用人 RBM15 启动子引物和人 ADGRL2 启动子引物,通过实时 PCR 对富集的 DNA 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。