Revision 6

#18665Store 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

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

23, 24

Source/Isotype:

Rabbit IgG

UniProt ID:

#P18847

Entrez-Gene Id:

467

Product Information

Product Usage Information

For optimal ChIP results, use 5 μl of antibody
and 10 μg of chromatin (approximately 4 × 10^6 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600
Chromatin IP 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

ATF-3 (E9J4N) Rabbit mAb recognizes endogenous levels of total ATF-3 protein.

Species Reactivity:

Human, Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant human ATF-3 protein.

Background

Activating transcription factor 3 (ATF-3) is a basic leucine zipper-type transcription factor belonging to the ATF/cAMP responsive element binding protein family. ATF-3 can form homodimers or heterodimers with other family members and depending on the cell types and promoter context, it can suppress or activate the transcription of its target genes (1,2). Expression of ATF-3 is induced by a variety of factors including cytokines, genotoxic agents, and physiological stress. For example, both metformin and high-density lipoprotein (HDL) can induce ATF-3 expression in macrophages (1,3-5). Research studies also indicate that ATF-3 can function as an oncogene or a tumor suppressor depending on the context (6,7).

  1. Hai, T. et al. (1999) Gene Expr 7, 321-35.
  2. Hai, T. and Hartman, M.G. (2001) Gene 273, 1-11.
  3. Hai, T. et al. (2010) Gene Expr 15, 1-11.
  4. De Nardo, D. et al. (2014) Nat Immunol 15, 152-60.
  5. Kim, J. et al. (2014) J Biol Chem 289, 23246-55.
  6. Yin, X. et al. (2008) Oncogene 27, 2118-27.
  7. Wang, Z. et al. (2014) Oncogene Advanced online publication, doi:10.1038/onc.2014.426.

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

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.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered 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 6
#18665

ATF-3 (E9J4N) Rabbit mAb

Western Blotting Image 1: ATF-3 (E9J4N) Rabbit mAb Expand Image
使用 ATF-3 (E9J4N) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对不同细胞系的提取物进行蛋白印迹分析。
Immunoprecipitation Image 1: ATF-3 (E9J4N) Rabbit mAb Expand Image
对 293T 细胞提取物 ATF-3 进行免疫沉淀。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 ATF-3 (E9J4N) Rabbit mAb。使用 ATF-3 (E9J4N) Rabbit mAb 进行蛋白印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Immunofluorescence Image 1: ATF-3 (E9J4N) Rabbit mAb Expand Image
使用 ATF-3 (E9J4N) Rabbit mAb(绿色)对 BeWo 细胞(左图,阳性)和 MCF7 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。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。
Flow Cytometry Image 1: ATF-3 (E9J4N) Rabbit mAb Expand Image
使用 ATF-3 (E9J4N) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对固定/透化的 NIH/3T3 细胞(蓝色,阴性)和 RAW 264.7 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。
Flow Cytometry Image 2: ATF-3 (E9J4N) Rabbit mAb Expand Image
使用 ATF-3 (E9J4N) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对固定/透化的 MCF7 细胞(蓝色,阴性)和 293T 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: ATF-3 (E9J4N) Rabbit mAb Expand Image
以 HCT 116 细胞的交联染色质与 ATF-3 (E9J4N) Rabbit mAb 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀分析(

使用 Simple ChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005)。使用 SimpleChIP® Human ATF-3 Exon 1 Primers #66427、human RUNX2 Intron 5 和 SimpleChIP® Human α Satellite Primers #4486,通过实时 PCR 对富集的 DNA 进行定量分析。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。