Revision 4

#62294Store 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, ChIP

REACTIVITY:

H M Mk

SENSITIVITY:

Endogenous

MW (kDa):

60

Source/Isotype:

Rabbit IgG

UniProt ID:

#P10276

Entrez-Gene Id:

5914

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
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

RARα (E6Z6K) Rabbit mAb recognizes endogenous levels of total RARα protein. This antibody weakly detects RARγ when it is overexpressed.

Species Reactivity:

Human, Mouse, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu220 of human RARα protein.

Background

Retinoids (vitamin A and its active retinoic acid derivatives) are non-steroid hormones that regulate cell proliferation, differentiation and apoptosis. Retinoic acid receptors (RARalpha, -beta and -gamma) and retinoid X receptors (RXRalpha, -beta and -gamma) are nuclear receptors that function as RAR-RXR heterodimers or RXR homodimers (1-2). In response to retinoid binding, these dimers control gene expression by binding to specific retinoic acid response elements, by recruiting cofactors and the transcriptional machinery, and by indirectly regulating chromatin structure. Finally, ligand binding and phosphorylation of RARalpha by JNK at Thr181, Ser445 and Ser461 controls the stability of RAR-RXR through the ubiquitin-proteasome pathway (3-4). At least four distinct genetic lesions affect RARalpha and result in acute promyelocytic leukemia (APL). The t(15;17) translocation that results in the PML-RARalpha fusion protein is responsible for more than 99% of APL cases, and the fusion protein inhibits PML-dependent apoptotic pathways in a dominant negative fashion. In addition PML-RARalpha inhibits transcription of retinoic acid target genes by recruiting co-repressors, attenuating myeloid differentiation (5-6).

  1. Mangelsdorf, D. J. et al. (1995) Cell 83, 835-839.
  2. Mangelsdorf, D.J. and Evans, R.M. (1995) Cell 83, 841-850.
  3. Bastien, J. and Rochette-Egly, C. (2004) Gene 328, 1-16.
  4. Srinivas, H. et al. (2005) Mol. Cell. Biol. 25, 1054-1069.
  5. de The, H. et al. (1990) Nature 347, 558-561.
  6. Slack, J.L. and Rusiniak, M.E. (2000) Ann. Hematol. 79, 227-238.

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 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 4
#62294

RARα (E6Z6K) Rabbit mAb

Western Blotting Image 1: RARα (E6Z6K) Rabbit mAb Expand Image
使用 RARα (E6Z6K) Rabbit mAb 对不同细胞系提取物进行的蛋白质印迹法分析。NB-4 细胞系包含 PML-RARα 融合蛋白。
Immunoprecipitation Image 1: RARα (E6Z6K) Rabbit mAb Expand Image
对来自 NB-4 细胞提取物的 RARα 和 PML-RARα 进行的免疫沉淀。泳道 1 为 10% 输入,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 RARα (E6Z6K) Rabbit mAb。使用 RARα (E6Z6K) Rabbit mAb 进行蛋白质印迹法分析。NB-4 细胞系包含 PML-RARα 融合。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。
Chromatin Immunoprecipitation Image 1: RARα (E6Z6K) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对 NB-4 细胞的交联染色质与 RARα (E6Z6K) Rabbit mAb #62294 或 Normal Rabbit IgG #2729 进行染色质免疫沉淀。使用 SimpleChIP® Human RUNX1 Exon 2 Primers #67254、人 GFI 启动子引物、人 GFI 外显子4引物和 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。