Overview

  • Product nameAnti-KMT2A / MLL antibody [mmN4.4]
    See all KMT2A / MLL primary antibodies
  • Description
    Mouse monoclonal [mmN4.4] to KMT2A / MLL
  • Tested applicationsSuitable for: IHC-P, Flow Cyt, IP, WB, ELISAmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant MBP fusion protein containing residues 161 - 356 of Human MLL.

  • Positive control
    • Human lung adenocarcinoma FFPE human tissue sections
  • General notesWe can conjugate this antibody to FITC for you (please see ab150234 for details).

Properties

Applications

Our Abpromise guarantee covers the use of ab32400 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IP Use at an assay dependent concentration.
WB Use a concentration of 10 µg/ml. Detects a band of approximately 432 kDa (predicted molecular weight: 432 kDa).
ELISA Use at an assay dependent concentration.

Target

  • FunctionHistone methyltransferase that plays an essential role in early development and hematopoiesis. Catalytic subunit of the MLL1/MLL complex, a multiprotein complex that mediates both methylation of 'Lys-4' of histone H3 (H3K4me) complex and acetylation of 'Lys-16' of histone H4 (H4K16ac). In the MLL1/MLL complex, it specifically mediates H3K4me, a specific tag for epigenetic transcriptional activation. Has weak methyltransferase activity by itself, and requires other component of the MLL1/MLL complex to obtain full methyltransferase activity. Has no activity toward histone H3 phosphorylated on 'Thr-3', less activity toward H3 dimethylated on 'Arg-8' or 'Lys-9', while it has higher activity toward H3 acetylated on 'Lys-9'. Required for transcriptional activation of HOXA9. Promotes PPP1R15A-induced apoptosis.
  • Tissue specificityHeart, lung, brain and T- and B-lymphocytes.
  • Involvement in diseaseNote=Chromosomal aberrations involving MLL are a cause of acute leukemias. Translocation t(1;11)(q21;q23) with MLLT11/AF1Q; translocation t(3;11)(p21;q23) with NCKIPSD/AF3p21; translocation t(3,11)(q25,q23) with GMPS; translocation t(4;11)(q21;q23) with AFF1/MLLT2/AF4; insertion ins(5;11)(q31;q13q23) with AFF4/AF5Q31; translocation t(5;11)(q12;q23) with AF5-alpha/CENPK; translocation t(6;11)(q27;q23) with MLLT4/AF6; translocation t(9;11)(p22;q23) with MLLT3/AF9; translocation t(10;11)(p11.2;q23) with ABI1; translocation t(10;11)(p12;q23) with MLLT10/AF10; t(11;15)(q23;q14) with CASC5 and ZFYVE19; translocation t(11;17)(q23;q21) with MLLT6/AF17; translocation t(11;19)(q23;p13.3) with ELL; translocation t(11;19)(q23;p13.3) with MLLT1/ENL; translocation t(11;19)(q23;p23) with GAS7; translocation t(X;11)(q13;q23) with FOXO4/AFX1. Translocation t(3;11)(q28;q23) with LPP. Translocation t(10;11)(q22;q23) with TET1. Translocation t(9;11)(q34;q23) with DAB2IP. Translocation t(4;11)(p12;q23) with FRYL. Fusion proteins MLL-MLLT1, MLL-MLLT3 and MLL-ELL interact with PPP1R15A and, on the contrary to unfused MLL, inhibit PPP1R15A-induced apoptosis.
    Note=A chromosomal aberration involving MLL may be a cause of chronic neutrophilic leukemia. Translocation t(4;11)(q21;q23) with SEPT11.
  • Sequence similaritiesBelongs to the histone-lysine methyltransferase family. TRX/MLL subfamily.
    Contains 3 A.T hook DNA-binding domains.
    Contains 1 bromo domain.
    Contains 1 CXXC-type zinc finger.
    Contains 1 FY-rich C-terminal domain.
    Contains 1 FY-rich N-terminal domain.
    Contains 3 PHD-type zinc fingers.
    Contains 1 post-SET domain.
    Contains 1 SET domain.
  • Domainthe 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
    The SET domain structure is atypical and is not in an optimal position to have methyltransferase activity. It requires other components of the MLL1/MLL complex, such as ASH2L or RBBP5, to order the active site and obtain optimal histone methyltransferase activity.
    The CXXC-type zinc finger binds bind to nonmethyl-CpG dinucleotides.
  • Post-translational
    modifications
    Proteolytic cleavage by TASP1 generates MLL cleavage product N320 and MLL cleavage product C180, which reassemble through a non-covalent association. 2 cleavage sites exist, cleavage site 1 (CS1) and cleavage site 2 (CS2), to generate MLL cleavage products N320 and C180. CS2 is the major site.
  • Cellular localizationNucleus and Nucleus. Localizes to a diffuse nuclear pattern when not associated with MLL cleavage product N320.
  • Information by UniProt
  • Database links
  • Alternative names
    • ALL-1 antibody
    • ALL1 antibody
    • C-terminal cleavage product of 180 kDa antibody
    • CXXC-type zinc finger protein 7 antibody
    • CXXC7 antibody
    • HRX antibody
    • HTRX1 antibody
    • KMT2A antibody
    • Lysine N-methyltransferase 2A antibody
    • Mll antibody
    • MLL cleavage product C180 antibody
    • MLL1 antibody
    • MLL1_HUMAN antibody
    • MLL1A antibody
    • N-terminal cleavage product of 320 kDa antibody
    • p180 antibody
    • p320 antibody
    • Trithorax-like protein antibody
    • TRX1 antibody
    • Zinc finger protein HRX antibody
    see all

Anti-KMT2A / MLL antibody [mmN4.4] images

  • Anti-KMT2A / MLL antibody [mmN4.4] (ab32400) at 10 µg/ml + Lysate from transfected cells overexpressing KMT2A/MLL

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size : 432 kDa
  • Overlay histogram showing HeLa cells stained with ab32400 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32400, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • IHC image of ab32400 staining in human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32400, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-KMT2A / MLL antibody [mmN4.4] (ab32400)

This product has been referenced in:
  • Gao Y  et al. Expression of p300 and CBP is associated with poor prognosis in small cell lung cancer. Int J Clin Exp Pathol 7:760-7 (2014). IHC-P ; Human . Read more (PubMed: 24551300) »
  • Chen P  et al. miR-9 is an essential oncogenic microRNA specifically overexpressed in mixed lineage leukemia-rearranged leukemia. Proc Natl Acad Sci U S A 110:11511-6 (2013). Read more (PubMed: 23798388) »

See all 5 Publications for this product

Product Wall

Abreviews
Application Western blot
Loading amount 90 µg
Gel Running Conditions Reduced Denaturing (6% gel)
Sample Human Cell lysate - whole cell (Human osteosarcoma U2OS cells)
Specification Human osteosarcoma U2OS cells
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted May 29 2013

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