Revision 4

#5528Store 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, W-S, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R Mk B Pg

SENSITIVITY:

Endogenous

MW (kDa):

90

Source/Isotype:

Rabbit IgG

UniProt ID:

#P51812

Entrez-Gene Id:

6197

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunohistochemistry (Paraffin) 1:800 - 1:3200
Immunofluorescence (Immunocytochemistry) 1:200 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200

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

RSK2 (D21B2) XP® Rabbit mAb recognizes endogenous levels of total RSK2 protein. This antibody does not cross-react with other RSK isoforms.

Species Reactivity:

Human, Mouse, Rat, Monkey, Bovine, Pig

Species predicted to react based on 100% sequence homology

Dog, Horse, Rabbit

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro686 of human RSK2 protein.

Background

The 90 kDa ribosomal S6 kinases (RSK1-4) are a family of widely expressed Ser/Thr kinases characterized by two nonidentical, functional kinase domains (1) and a carboxy-terminal docking site for extracellular signal-regulated kinases (ERKs) (2). Several sites both within and outside of the RSK kinase domain, including Ser380, Thr359, Ser363, and Thr573, are important for kinase activation (3). RSK1-3 are activated via coordinated phosphorylation by MAPKs, autophosphorylation, and phosphoinositide-3-OH kinase (PI3K) in response to many growth factors, polypeptide hormones, and neurotransmitters (3).

Stimulation by various growth factors leads to activation of RSK2, which is a critical downstream effector kinase in several pathways. EGF stimulation leads to phosphorylation of CREB at Ser133 and phosphorylation of histone H3 in vivo by RSK2 (4,5). RSK2 phosphorylation of p53 may help regulate chromatin structure and cell cycle (6). RSK2 is prominently expressed in the brain and is essential for cognitive function and learning. During development, RSK2 regulates the differentiation of osteoblasts and skeletal muscle cells (7,8). Mutations in the corresponding gene are associated with Coffin-Lowry syndrome (CLS), an X-linked disorder characterized by mental retardation and the presence of characteristic facial anomalies (9).

  1. Fisher, T.L. and Blenis, J. (1996) Mol Cell Biol 16, 1212-9.
  2. Smith, J.A. et al. (1999) J Biol Chem 274, 2893-8.
  3. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
  4. De Cesare, D. et al. (1998) Proc Natl Acad Sci U S A 95, 12202-7.
  5. Sassone-Corsi, P. et al. (1999) Science 285, 886-91.
  6. Cho, Y.Y. et al. (2005) Cancer Res 65, 3596-603.
  7. Yang, X. et al. (2004) Cell 117, 387-98.
  8. Cho, Y.Y. et al. (2007) J Biol Chem 282, 8380-92.
  9. Delaunoy, J.P. et al. (2006) Clin Genet 70, 161-6.

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 W-S: Simple Western™ IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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.
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
#5528

RSK2 (D21B2) XP® Rabbit mAb

Western Blotting Image 1: RSK2 (D21B2) XP® Rabbit mAb Expand Image
使用 RSK2 (D21B2) XP® 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: RSK2 (D21B2) XP® Rabbit mAb Expand Image
使用 RSK1 (D6D5) Rabbit mAb #8408(左小图)、RSK2 (D21B2) XP® Rabbit mAb #5528(中间小图)或 GST (91G1) Rabbit mAb #2625(右小图),对重组带有 GST 标记的 RSK1、RSK2 和 RSK3(各 2 μg)进行蛋白质印迹分析。
Western Blotting Image 1: RSK2 (D21B2) XP® Rabbit mAb Expand Image

Simple Western™ analysis of lysates (0.1 mg/mL) from COS-7 untreated cells using RSK2 (D21B2) XP® Rabbit mAb #5528. 虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的单一靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。

Immunohistochemistry Image 1: RSK2 (D21B2) XP® Rabbit mAb Expand Image
使用 RSK2 (D21B2) XP® Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: RSK2 (D21B2) XP® Rabbit mAb Expand Image
使用 RSK2 (D21B2) XP® Rabbit mAb 对石蜡包埋的人肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: RSK2 (D21B2) XP® Rabbit mAb Expand Image
使用 RSK2 (D21B2) XP® Rabbit mAb 对石蜡包埋的人淋巴瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 4: RSK2 (D21B2) XP® Rabbit mAb Expand Image
在有对照肽(左图)或抗原特异性肽(右图)的情况下,使用 RSK2 (D21B2) XP® Rabbit mAb 对石蜡包埋的人卵巢浆液性腺癌细胞进行免疫组织化学分析。
Immunofluorescence Image 1: RSK2 (D21B2) XP® Rabbit mAb Expand Image
使用 RSK2 (D21B2) XP® Rabbit mAb(绿色),对未经处理(左图)或已经亚砷酸钠(500 uM,1 小时;右图)处理的 MCF7 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。RSK2 在经亚砷酸盐处理细胞中“应激颗粒”的位置与 Eisinger-Mathason 等人看到的位置非常相似。(分子细胞,卷 1, 2008.09.5,第 722-736 页)。
Immunofluorescence Image 2: RSK2 (D21B2) XP® Rabbit mAb Expand Image
使用 RSK2 (D21B2) XP® Rabbit mAb(绿色),对未经处理(左图)或已经胰岛素、EGF 和血清(右图)处理的 MCF7 细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: RSK2 (D21B2) XP® Rabbit mAb Expand Image
以 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(红色)作为对照,使用 RSK2 (D21B2) XP® Rabbit mAb(蓝色)对 Hela 细胞进行流式细胞分析。