Revision 1

#3085Store 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 R

SENSITIVITY:

Endogenous

MW (kDa):

53

Source/Isotype:

Rabbit IgG

UniProt ID:

#P19793

Entrez-Gene Id:

6256

Product Information

Product Usage Information

For optimal ChIP 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.

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

RXRα (D6H10) Rabbit mAb recognizes endogenous levels of total RXRα protein. This antibody does not cross-react with either RXRβ or RXRγ.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human RXRα protein.

Background

The human retinoid X receptors (RXRs) are encoded by three distinct genes (RXRα, RXRβ, and RXRγ) and bind selectively and with high affinity to the vitamin A derivative, 9-cis-retinoic acid. RXRs are type-II nuclear hormone receptors that are largely localized to the nuclear compartment independent of ligand binding. Nuclear RXRs form heterodimers with nuclear hormone receptor subfamily 1 proteins, including thyroid hormone receptor, retinoic acid receptors, vitamin D receptor, peroxisome proliferator-activated receptors, liver X receptors, and farnesoid X receptor (1). Since RXRs heterodimerize with multiple nuclear hormone receptors, they play a central role in transcriptional control of numerous hormonal signaling pathways by binding to cis-acting response elements in the promoter/enhancer region of target genes (2).
Retinoid X receptor α (RXRα) is the founding RXR family member and is predominantly expressed in the liver, kidney, epidermis, intestine, and a variety of tissues (2-4). Knockout of the murine rxrα gene results in embryonic lethality tentatively due to myocardial hypoplasia, which demonstrates the importance of RXRα in retinoid signaling in vivo (5,6). Biochemical evidence suggests that RXRα transcriptional activity is post-translationally regulated through the Ras-Raf-MAPK signaling cascade. MAPK-dependent phosphorylation of RXRα directly abrogates the ability of RXRα to associate with nuclear receptor coactivators (7).

  1. Gronemeyer, H. et al. (2004) Nat Rev Drug Discov 3, 950-64.
  2. Mangelsdorf, D.J. et al. (1992) Genes Dev 6, 329-44.
  3. Mangelsdorf, D.J. et al. (1990) Nature 345, 224-9.
  4. Dollé, P. et al. (1994) Mech Dev 45, 91-104.
  5. Kastner, P. et al. (1994) Cell 78, 987-1003.
  6. Sucov, H.M. et al. (1994) Genes Dev 8, 1007-18.
  7. Macoritto, M. et al. (2008) J Biol Chem 283, 4943-56.

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 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.
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 1
#3085

RXRα (D6H10) Rabbit mAb

Western Blotting Image 1: RXRα (D6H10) Rabbit mAb Expand Image
使用 RXRα (D6H10) Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
Western Blotting Image 2: RXRα (D6H10) Rabbit mAb Expand Image
使用 RXRα (D6H10) Rabbit mAb (上图)和 DYKDDDDK Tag Antibody (Binds to same epitope as Sigma's Anti-FLAG® M2 Antibody) #2368(下图),对转染空载 (-) 或转染能编码全长人 RXRα (hRXRα; +)、RXRβ (hRXRβ; +) 或 RXRγ (hRXRγ; +) 的带有 Myc/DDK 标签的 cDNA 表达载体的 293T 细胞的提取物进行蛋白质印迹分析。
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Chromatin Immunoprecipitation Image 1: RXRα (D6H10) Rabbit mAb Expand Image
使用 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005 对经维生素 D (10 nM) 处理 3 小时的 LS-180 细胞的交联染色质与 RXRα (D6H10) Rabbit mAb 或 Normal Rabbit IgG #2729 一起进行染色质免疫沉淀分析。使用 SimpleChIP® Human c-Fos Upstream Primers #25661、human UCA1 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。