Revision 8

#53528Store 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

SENSITIVITY:

Endogenous

MW (kDa):

50

Source/Isotype:

Rabbit IgG

UniProt ID:

#P55317

Entrez-Gene Id:

3169

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 1:50
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:3200
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600
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

FoxA1/HNF3α (E7E8W) Rabbit mAb recognizes endogenous levels of total FoxA1/HNF3α protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human FoxA1/HNF3α protein.

Background

Forkhead box protein A1 (FoxA1, HNF3α) is a transcription factor required for the development of endoderm-derived organs, such as liver, lung, and prostate (1). FoxA1 functions as a pioneer factor that is recruited primarily to the distant enhancers to change chromatin structure for transcription in a cell type-specific manner (2,3). The FoxA1 transcription factor is implicated in various diseases, playing a role in hormone-dependent disorders, such as breast and prostate cancers (4). The treatment of relapsing-remitting multiple sclerosis patients with IFN-β results in FoxA1-induced stimulation of a novel population of FoxA1(+) regulatory T cells, suggesting a possible immunosuppressive role for FoxA1 (5).

  1. Friedman, J.R. and Kaestner, K.H. (2006) Cell Mol Life Sci 63, 2317-28.
  2. Lupien, M. et al. (2008) Cell 132, 958-70.
  3. Wang, A. et al. (2015) Cell Stem Cell 16, 386-99.
  4. Augello, M.A. et al. (2011) EMBO J 30, 3885-94.
  5. Liu, Y. et al. (2014) Nat Med 20, 272-82.

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 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 8
#53528

FoxA1/HNF3α (E7E8W) Rabbit mAb

Western Blotting Image 1: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 FoxA1/HNF3α (E7E8W) 兔单克隆抗体(上图)和 β-肌动蛋白 (D6A8) 兔单克隆抗体 #8457(下图)对不同细胞系的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
对 Hep G2 提取物 FoxA1/HNF3α 进行免疫沉淀。泳道 1 为 10% 输入,泳道 2 为兔 (DA1E) 兔单克隆抗体 IgG XP® 同型对照 #3900,泳道 3 为 FoxA1/HNF3α (E7E8W) 单克隆抗体。使用 FoxA1/HNF3α (E7E8W) Rabbit mAb 进行蛋白质印迹法分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Immunofluorescence Image 1: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 FOXA1 (E7E8W) 单克隆抗体(绿色)对 MCF7 细胞(左图,阳性)和 SK-MEL-5 细胞(右图,阴性)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: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 FoxA1/HNF3α (E7E8W) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对固定和透化的 Daudi 细胞(蓝色,阴性)和 MCF7 细胞(绿色,阳性)进行流式细胞分析。Anti-rabbit IgG F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
Chromatin Immunoprecipitation Image 1: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 LNCaP 细胞的交联染色质和 FoxA1/HNF3α (E7E8W) 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 KLK3 内结合。
Chromatin Immunoprecipitation Image 2: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 LNCaP 细胞的交联染色质和 FoxA1/HNF3α (E7E8W) 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 19(上图)内的结合,包括 FoxA1/HNF3α 的已知靶基因 KLK3(下图)。
Chromatin Immunoprecipitation Image 3: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对未处理的 LNCaP 细胞的交联染色质与 FoxA1/HNF3α (E7E8W) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀。使用 human SLUG exon 3 primers、human PSA enhancer primers、SimpleChIP® Human KLK3 Promoter Primers #32784 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。
CUT and RUN Image 1: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 LNCaP 细胞和 FoxA1/HNF3α (E7E8W) Rabbit mAb 进行 CUT&RUN 分析。使用 DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示了 FoxA1 的已知靶标基因 RABL3 内的结合(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 2: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对 LNCaP 细胞和 FoxA1/HNF3α (E7E8W) Rabbit mAb 进行 CUT&RUN 分析。使用 DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795 制备 DNA 库。该图显示了在染色体 3(上图)内的结合,包括 FoxA1 的已知靶基因 RABL3(下图)(参见包含 CUT&RUN-qPCR 数据的其他结果图)。
CUT and RUN Image 3: FoxA1/HNF3α (E7E8W) Rabbit mAb Expand Image
使用 CUT&RUN Assay Kit #86652 对€‚ LNCaP 细胞和 FoxA1/HNF3α (E7E8W) Rabbit mAb 或 Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362 进行 CUT&RUN 检测。使用 human RABL3 promoter primers、human CCDC190 promoter primer 和 SimpleChIP® Human α Satellite Repeat Primers #4486,通过实时定量 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。