Revision 1

#64733Store 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 Mk

SENSITIVITY:

Endogenous

MW (kDa):

80

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9NVC6

Entrez-Gene Id:

9440

Product Information

Product Usage Information

For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:200
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

MED17 (E3V6Y) Rabbit mAb recognizes endogenous levels of total MED17 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly272 of human MED17 protein.

Background

The mammalian Mediator complex is a multi-subunit protein complex that acts to relay signals from transcription factors to RNA polymerase II (Pol II) and the basal transcription machinery. In other words, Mediator acts as a centralized hub or integrator for transcriptional regulation (reviewed in 1,2). Multiple studies have shown that Mediator functions include epigenetic regulation, transcriptional elongation, transcription termination, mRNA processing, non-coding RNA activation, and super-enhancer formation. Through these functions, Mediator contributes to the regulation of many biological processes, including insulin signaling (3), NF-κB-dependent signaling (4), stem cell pluripotency and self-renewal (5,6), and proliferation of colon cancer cells (7,8). The Mediator complex consists of 31 protein subunits and can be divided into four distinct modules: the head, middle, tail, and CDK8 kinase module. The CDK8 kinase module contains the CDK8, cyclin C, MED12, and MED13 subunits and is thought to act as a molecular switch, phosphorylating the C-terminal domain (CTD) of Rpb1, the large subunit of Pol II, to inhibit Pol II recruitment and transcription initiation (9,10).

MED17, along with MED6, 8, 11, 18, 20, and 22, make up the head module of the Mediator complex (11). MED17 is critical for head module assembly as it acts as a scaffold to recruit MED11 and MED22 (12). Phosphorylation of MED17 has been shown as a regulatory mechanism in lipogenesis and NF-κB signaling (13,14).

  1. Malik, S. and Roeder, R.G. (2005) Trends Biochem Sci 30, 256-63.
  2. Soutourina, J. (2018) Nat Rev Mol Cell Biol 19, 262-274.
  3. Wang, W. et al. (2009) Dev Cell 16, 764-71.
  4. van Essen, D. et al. (2009) PLoS Biol 7, e73.
  5. Tutter, A.V. et al. (2009) J Biol Chem 284, 3709-18.
  6. Varelas, X. et al. (2008) Nat Cell Biol 10, 837-48.
  7. Firestein, R. et al. (2008) Nature 455, 547-51.
  8. Morris, E.J. et al. (2008) Nature 455, 552-6.
  9. Knuesel, M.T. et al. (2009) Mol Cell Biol 29, 650-61.
  10. Knuesel, M.T. et al. (2009) Genes Dev 23, 439-51.
  11. Takagi, Y. et al. (2006) Mol Cell 23, 355-64.
  12. Imasaki, T. et al. (2011) Nature 475, 240-3.
  13. Viscarra, J.A. et al. (2017) Sci Signal 10, .
  14. Mashukova, A. et al. (2021) Mol Biol Cell 32, 690-702.

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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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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
#64733

MED17 (E3V6Y) Rabbit mAb

Western Blotting Image 1: MED17 (E3V6Y) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using MED17 (E3V6Y) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457.
Western Blotting Image 2: MED17 (E3V6Y) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human MED17 (MED17-Myc/DDK; +), using MED17 (E3V6Y) Rabbit mAb (upper) or DYKDDDDK Tag (D6W5B) Rabbit mAb #14793 (lower).
Immunoprecipitation Image 1: MED17 (E3V6Y) Rabbit mAb Expand Image
Immunoprecipitation of MED17 protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MED17 (E3V6Y) Rabbit mAb. Western blot analysis was performed using MED17 (E3V6Y) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Chromatin Immunoprecipitation Image 1: MED17 (E3V6Y) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells, heatshocked at 42°C for 1 hr, and either MED17 (E3V6Y) Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR, using SimpleChIP® Human HSP70 Intron 1 Primers #14023, SimpleChIP® Human c-Myc Intron 1 Primers #14905, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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。