Revision 1
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, IF-IC

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

180

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q03164

Entrez-Gene Id:

4297

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:200

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

MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) recognizes endogenous levels of total MLL1-C protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human MLL1 protein.

Background

The Set1 histone methyltransferase protein was first identified in yeast as part of the Set1/COMPASS histone methyltransferase complex, which methylates histone H3 at Lys4 and functions as a transcriptional co-activator (1). While yeast contain only one known Set1 protein, mammals contain six Set1-related proteins: SET1A, SET1B, MLL1, MLL2, MLL3, and MLL4, all of which assemble into COMPASS-like complexes and methylate histone H3 at Lys4 (2,3). These Set1-related proteins are each found in distinct protein complexes, all of which share the common subunits WDR5, RBBP5, ASH2L, CXXC1 and DPY30, which are required for proper complex assembly and modulation of histone methyltransferase activity (2-6). MLL1 and MLL2 complexes contain the additional protein subunit, menin (6).

MLL1 functions as a master regulator of both embryogenesis and hematopoiesis, and is required for proper expression of Hox genes (7,8). MLL1 is a large, approximately 4000 amino acid, protein that is cleaved by the taspase 1 threonine endopeptidase to form N-terminal (MLL1-N) and C-terminal MLL1 (MLL1-C) fragments, both of which are subunits of the functional MLL1/COMPASS complex (9,10). MLL1-N, MLL1-C, WDR5, RBBP5 and ASH2L define the core catalytic component of the MLL1/COMPASS complex, which is recruited to target genes and methylates histone H3 lysine 4 to regulate transcriptional initiation (11). At least 60 different MLL1 translocation partners have been molecularly characterized and associated with various hematological malignancies. The most common translocation partners include AF4, AF9, ENL, AF10, ELL and AF6 (8,12,13). With the exception of AF6, all of these partners are nuclear proteins that function to positively regulate transcriptional elongation. AF4, AF9 and ENL are all components of the super elongation complex (SEC), while AF4, AF9, AF10 and ENL all interact with the histone H3 lysine 79 methyltransferase DOT1L. Many MLL1 target genes are normally regulated by promoter-proximal pausing, with the release of RNA polymerase and transcriptional elongation occurring in response to proper stimuli (14). The association of MLL1 translocation partners with SEC and DOT1L suggest that MLL1-fusion proteins may function to sustain specific gene expression programs by constitutively activating transcriptional elongation.

  1. Miller, T. et al. (2001) Proc Natl Acad Sci U S A 98, 12902-7.
  2. Shilatifard, A. (2008) Curr Opin Cell Biol 20, 341-8.
  3. Tenney, K. and Shilatifard, A. (2005) J Cell Biochem 95, 429-36.
  4. Lee, J.H. and Skalnik, D.G. (2005) J Biol Chem 280, 41725-31.
  5. Lee, J.H. et al. (2007) J Biol Chem 282, 13419-28.
  6. Hughes, C.M. et al. (2004) Mol Cell 13, 587-97.
  7. Eissenberg, J.C. and Shilatifard, A. (2010) Dev Biol 339, 240-9.
  8. Smith, E. et al. (2011) Genes Dev 25, 661-72.
  9. Takeda, S. et al. (2006) Genes Dev 20, 2397-409.
  10. Yokoyama, A. et al. (2002) Blood 100, 3710-8.
  11. Dou, Y. et al. (2006) Nat Struct Mol Biol 13, 713-9.
  12. Yip, B.H. and So, C.W. (2013) Exp Biol Med (Maywood) 238, 315-23.
  13. Neff, T. and Armstrong, S.A. (2013) Blood 121, 4847-53.
  14. Wang, P. et al. (2009) Mol Cell Biol 29, 6074-85.

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 IF-IC: Immunofluorescence (Immunocytochemistry)

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.
XP 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.

使用に関する制限

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Revision 1
#14197

MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen)

Western Blotting Image 1: MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) Expand Image
Western blot analysis of extracts from MLL1 wild-type (WT) and knockout (KO) mouse embryonic fibroblasts (MEF) using MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). MLL1 WT and KO MEF were kindly provided by Dr. Ali Shilatifard of Northwestern University.
Western Blotting Image 2: MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) Expand Image
Western blot analysis of extracts from various cell lines using MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen).
No image available
Immunofluorescence Image 1: MLL1 (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) Expand Image
Confocal immunofluorescent analysis of MEF wild-type (left, positive) and MEF MLL (-/-) (right, negative) cells using MLL (D6G8N) Rabbit mAb (Carboxy-terminal Antigen) (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). MLL1 WT and KO MEF were kindly provided by Dr. Ali Shilatifard of Northwestern University.