Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

ELL (D7N6U) Rabbit mAb #14468

No. Size Price
14468S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
14468 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 70 Rabbit IgG
IP 1:100
IF-IC 1:1600
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Bovine, Dog, Horse,

Specificity / Sensitivity

ELL (D7N6U) Rabbit mAb recognizes endogenous levels of total ELL protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp399 of human ELL protein.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using ELL (D7N6U) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using ELL (D7N6U) Rabbit mAb.

IP

IP

Immunoprecipitation of ELL from 293T cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ELL (D7N6U) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using ELL (D7N6U) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293T cells and either 5 µl of ELL (D7N6U) Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human RNU2-1 Promoter Primers #14084 and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

The super elongation complex (SEC) plays a critical role in regulating RNA polymerase II (RNAPII) transcription elongation (1). The SEC is composed of AFF4, AFF1/AF4, MLLT3/AF9, and MLLT1/ENL proteins. The pathogenesis of mixed lineage leukemia is often associated with translocations of the SEC subunits joined to the histone H3 Lys4 methyltransferase mixed lineage leukemia (MLL) gene (1-4). The SEC has been found to contain RNAPII elongation factors eleven-nineteen lysine-rich leukemia (ELL), ELL2, and ELL3, along with the associated factors EAF1 and EAF2, which can increase the catalytic rate of RNAPII transcription in vitro, (1,2,5-7). The SEC positive transcription elongation factor b (P-TEFb) phosphorylates the carboxy-terminal domain within the largest subunit of RNAP II at Ser2 of the heptapeptide repeat. The SEC negative transcription elongation factors, DRB-induced stimulating factor (DSIF) and negative elongation factor (NELF), signal the transition from transcription initiation and pausing to productive transcription elongation (2,8-10). The chromosomal translocation of MLL with the members of the SEC leads to SEC recruitment to MLL regulated genes, such as the highly developmentally regulated Hox genes, implicating the misregulation and overexpression of these genes as underlying contributors to leukemogenesis (1,2,9,11).

  1. Mohan, M. et al. (2010) Nat Rev Cancer 10, 721-8.
  2. Lin, C. et al. (2010) Mol Cell 37, 429-37.
  3. Drexler, H.G. et al. (2004) Leukemia 18, 227-32.
  4. Smith, E. et al. (2011) Genes Dev 25, 661-72.
  5. Shilatifard, A. et al. (1996) Science 271, 1873-6.
  6. Shilatifard, A. et al. (1997) Proc Natl Acad Sci U S A 94, 3639-43.
  7. Miller, T. et al. (2000) J Biol Chem 275, 32052-6.
  8. Lin, C. et al. (2011) Genes Dev 25, 1486-98.
  9. Yokoyama, A. et al. (2010) Cancer Cell 17, 198-212.
  10. Cho, S. et al. (2010) Cell Cycle 9, 1697-705.
  11. Shah, N. and Sukumar, S. (2010) Nat Rev Cancer 10, 361-71.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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