WB, ChIP, ChIP-seq, C&R
H
Endogenous
80
Rabbit IgG
#Q03111
4298
Product Information
Product Usage Information
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Chromatin IP | 1:50 |
| Chromatin IP-seq | 1:50 |
| CUT&RUN | 1:50 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala343 of human MLLT1/ENL protein.
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).
MLL translocated to 1/eleven-nineteen-leukemia (MLLT1/ENL) is also found as part of the histone H3 Lys79 methyltransferase disruptor of telomeric silencing-like (Dot1L) complex that has been suggested to play a role in transcription elongation. This complex regulates the expression of genes, such as the Wnt-signaling pathway target genes that control cell proliferation and differentiation during development (12,13).
- Mohan, M. et al. (2010) Nat Rev Cancer 10, 721-8.
- Lin, C. et al. (2010) Mol Cell 37, 429-37.
- Drexler, H.G. et al. (2004) Leukemia 18, 227-32.
- Smith, E. et al. (2011) Genes Dev 25, 661-72.
- Shilatifard, A. et al. (1996) Science 271, 1873-6.
- Shilatifard, A. et al. (1997) Proc Natl Acad Sci U S A 94, 3639-43.
- Miller, T. et al. (2000) J Biol Chem 275, 32052-6.
- Lin, C. et al. (2011) Genes Dev 25, 1486-98.
- Yokoyama, A. et al. (2010) Cancer Cell 17, 198-212.
- Cho, S. et al. (2010) Cell Cycle 9, 1697-705.
- Shah, N. and Sukumar, S. (2010) Nat Rev Cancer 10, 361-71.
- Mohan, M. et al. (2010) Genes Dev 24, 574-89.
- Nguyen, A.T. and Zhang, Y. (2011) Genes Dev 25, 1345-58.
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 ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq C&R: CUT&RUN
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
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