Revision 5
#14893
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, ChIP, ChIP-seq, C&R
Reactivity:
H
Sensitivity:
Endogenous
MW (kDa):
80
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q03111
Entrez-Gene Id:
4298
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
MLLT1/ENL (D9M4B) Rabbit mAb recognizes endogenous levels of total MLLT1/ENL protein.
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).
Background References
- 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
W: Western Blotting ChIP: Chromatin IP 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 G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SimpleChIP 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.
Limited Uses
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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 5
#14893
MLLT1/ENL (D9M4B) Rabbit mAb
Western blot analysis of extracts from various cell lines using MLLT1/ENL (D9M4B) Rabbit mAb.
Chromatin immunoprecipitations were performed with cross-linked chromatin from MV-4-11 cells and MLLT1/ENL (D9M4B) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across MEIS1, a known target gene of MLLT1/ENL (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Chromatin immunoprecipitations were performed with cross-linked chromatin from MV-4-11 cells and MLLT1/ENL (D9M4B) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 2 (upper), including MEIS1 (lower), a known target gene of MLLT1/ENL (see additional figure containing ChIP-qPCR data).
Revision 5
#14893
MLLT1/ENL (D9M4B) Rabbit mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from MV-4-11 cells and either MLLT1/ENL (D9M4B) Rabbit mAb or 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 HoxA9 Promoter Primers #14909, human MEIS1 promoter primers, 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.
CUT&RUN was performed with MV-4-11 cells and MLLT1/ENL (D9M4B) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across MEIS1, a known target gene of MLLT1 (see additional figure containing CUT&RUN-qPCR data).
CUT&RUN was performed with MV-4-11 cells and MLLT1/ENL (D9M4B) Rabbit mAb, using CUT&RUN Assay Kit #86652. DNA library was prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 2 (upper), including MEIS1 (lower), a known target gene of MLLT1 (see additional figure containing CUT&RUN-qPCR data).
Revision 5
#14893
MLLT1/ENL (D9M4B) Rabbit mAb
CUT&RUN was performed with MV-4-11 cells and either MLLT1/ENL (D9M4B) Rabbit mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human HoxA9 Promoter Primers #14909, human MEIS1 promoters, 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.