Revision 4

#2316Store 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, IHC-P, IF-IC, FC-FP, eCLIP

REACTIVITY:

H M R Hm Mk B Dg

SENSITIVITY:

Endogenous

MW (kDa):

42, 55

Source/Isotype:

Rabbit 

UniProt ID:

#P50750

Entrez-Gene Id:

1025

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:50 - 1:100
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400
eCLIP 1:200
For more information about the RBP-eCLIP service please visit Eclipsebio.

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

CDK9 (C12F7) Rabbit mAb detects endogenous levels of total CDK9 protein, both 42 kDa and 55 kDa isoforms.

Species Reactivity:

Human, Mouse, Rat, Hamster, Monkey, Bovine, Dog

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human CDK9.

Background

P-TEFb is a general transcription factor that regulates transcription elongation through phosphorylation of the C-terminal tail domain (CTD) of RNA polymerase II (RNAP II). The P-TEFb complex is composed of a catalytic subunit, CDK9, and its regulatory cyclin partner, which can be cyclin T1, T2a, T2b or K (reviewed in 1,2). P-TEFb is recruited by the HIV Tat protein to allow transcriptional elongation, and subsequent replication of the viral genome. Inhibition of P-TEFb function therefore has potential for HIV therapy. CDK9 exists as two isoforms, an abundant 42 kDa isoform, and a less abundant 55 kDa isoform, which contains an amino-terminal extension (3). The two forms likely have distinct purposes based on differential expression during lymphocyte activation (4,5) and on their localization within the nucleus (5).Cyclin dependent kinases (CDKs) are activated in part by cyclin binding and by phosphorylation of a conserved threonine in the T-loop domain. Phosphorylation of CDK9 at the T-loop Thr186 by an unidentified nuclear kinase may be important in P-TEFb activation (6) and regulation of HIV transcription (7). Acetylation of CDK9 at Lys44 affects its ability to phosphorylate the RNAPII CTD (8).

  1. Rice, A.P. and Herrmann, C.H. (2003) Curr HIV Res 1, 395-404.
  2. De Falco, G. and Giordano, A. (2002) Cancer Biol Ther 1, 342-7.
  3. Shore, S.M. et al. (2003) Gene 307, 175-82.
  4. Shore, S.M. et al. (2005) Gene 350, 51-8.
  5. Liu, H. and Herrmann, C.H. (2005) J Cell Physiol 203, 251-60.
  6. Chen, R. et al. (2004) J Biol Chem 279, 4153-60.
  7. Ammosova, T. et al. (2005) Retrovirology 2, 47.
  8. Fu, J. et al. (2007) Mol Cell Biol 27, 4641-51.

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 IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) eCLIP: eCLIP

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.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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
#2316

CDK9 (C12F7) Rabbit mAb

Western Blotting Image 1: CDK9 (C12F7) Rabbit mAb Expand Image
使用 CDK9 (C12F7) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: CDK9 (C12F7) Rabbit mAb Expand Image
使用 CDK9 (C12F7) Rabbit mAb 对 HeLa 细胞的 CDK9 进行免疫沉淀。使用相同的抗体进行蛋白质印迹检测。泳道 1 是 5% 输入对照。
Immunohistochemistry Image 1: CDK9 (C12F7) Rabbit mAb Expand Image
在对照肽(左)或抗原特异性肽(右)存在的情况下,使用 CDK9 (C12F7) Rabbit mAb 对石蜡包埋的人乳腺癌进行免疫组织化学分析。
Immunohistochemistry Image 2: CDK9 (C12F7) Rabbit mAb Expand Image
使用 CDK9 (C12F7) Rabbit mAb 对石蜡包埋的 K7M2 小鼠同基因肿瘤进行免疫组织化学分析。
Immunofluorescence Image 1: CDK9 (C12F7) Rabbit mAb Expand Image
使用 CDK9 (C12F7) Rabbit mAb(绿色)对 Hela 细胞进行共聚焦免疫荧光分析。肌动蛋白丝可用 DY-555 Phalloidin(红色)标记。
Flow Cytometry Image 1: CDK9 (C12F7) Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 CDK9 (C12F7) Rabbit mAb(实线)对 Hela 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
eCLIP Image 1: CDK9 (C12F7) Rabbit mAb Expand Image
Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from Hep G2 cells and CDK9 (C12F7) Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. The figure shows binding across the PUM2 transcript. 数据由 Gene Yeo 博士的实验室友情提供,并经许可使用。