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

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

320

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9UMN6

Entrez-Gene Id:

9757

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100

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

MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen) recognizes endogenous levels of total MLL2/KMT2B protein. This antibody detects the Taspase 1-cleaved 320 kDa N-terminal MLL2/KMT2B protein (MLL2/KMT2B-N) and the full-length 400 kDa MLL2/KMT2B protein (MLL2/KMT2B-FL).

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala480 of human MLL2/KMT2B 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).
MLL2, also known as histone-lysine N-methyltransferase 2B (KMT2B), functions to activate gene expression by mediating tri-methylation of histone H3 lysine 4 at the promoters of genes involved in embryogenesis and hematopoiesis, and is required for histone H3 lysine 4 tri-methylation at bivalent promoters in embryonic stem cells (7). Like MLL1, MLL2 is a large protein made up of approximately 2,700 amino acids that is cleaved by the Taspase 1 threonine endopeptidase to form N-terminal (MLL2-N) and C-terminal (MLL2-C) fragments, both of which are subunits of the functional MLL2/COMPASS complex. MLL2-N, MLL2-C, WDR5, RBBP5, and ASH2L define the core catalytic component of the MLL2/COMPASS complex, which is recruited to target genes to regulate transcription. MLL1 gene translocations are often associated with various hematological malignancies and thought to be a driving component of these types of leukemia. MLL2 is required for memory formation, proper glucose homeostasis, and cardiac lineage differentiation of mouse embryonic stem cells (8-11). A recent study has shown that MLL2 is required for survival of MLL-AF9-transformed cells, implicating MLL2 as a potential modulator of MLL1-rearranged leukemias (12). Mutations in MLL2 cause complex early-onset dystonia, and overexpression of MLL2 is associated with gastrointestinal diffuse large B-cell lymphoma (13,14).

  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. Denissov, S. et al. (2014) Development 141, 526-37.
  8. Kerimoglu, C. et al. (2017) Cell Rep 20, 538-48.
  9. Kerimoglu, C. et al. (2013) J Neurosci 33, 3452-64.
  10. Goldsworthy, M. et al. (2013) PLoS One 8, e61870.
  11. Wan, X. et al. (2014) Stem Cell Rev 10, 643-52.
  12. Chen, Y. et al. (2017) Cancer Cell 31, 755-770.e6.
  13. Meyer, E. et al. (2017) Nat Genet 49, 223-37.
  14. Ye, H. et al. (2015) Int J Clin Exp Pathol 8, 13043-50.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

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
#47097

MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen)

Western Blotting Image 1: MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen) Expand Image
Western blot analysis of extracts from control 293T cells (lane 1) or MLL2/KMT2B knockout 293T cells (lane 2) using MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen) (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The absence of signal in the MLL2/KMT2B knockout 293T cells confirms specificity of the antibody for MLL2/KMT2B.
Western Blotting Image 2: MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen) Expand Image
Western blot analysis of extracts from NCCIT, H-4-II-E, and COS-7 cells using MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen).
Immunoprecipitation Image 1: MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen) Expand Image
Immunoprecipitation of MLL2/KMT2B protein from NCCIT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen). Western blot analysis was performed using MLL2/KMT2B (E3M1V) Rabbit mAb (Amino-terminal Antigen). Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.