Mouse Monoclonal KMT2A / MLL antibody. Suitable for IHC-P, Flow Cyt (Intra), WB and reacts with Human, Transfected cell lysate samples. Cited in 8 publications.
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
IHC-P | Flow Cyt (Intra) | WB | |
---|---|---|---|
Human | Tested | Tested | Expected |
Transfected cell lysate | Not recommended | Not recommended | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info 5 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Transfected cell lysate | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info 1 µg for 106 Cells | Notes ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Transfected cell lysate | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Transfected cell lysate | Dilution info 10 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
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Histone methyltransferase that plays an essential role in early development and hematopoiesis (PubMed:12453419, PubMed:15960975, PubMed:19187761, PubMed:19556245, PubMed:20677832, PubMed:21220120, PubMed:26886794). 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) (PubMed:12453419, PubMed:15960975, PubMed:19187761, PubMed:19556245, PubMed:20677832, PubMed:21220120, PubMed:24235145, PubMed:26886794). Catalyzes methyl group transfer from S-adenosyl-L-methionine to the epsilon-amino group of 'Lys-4' of histone H3 (H3K4) via a non-processive mechanism. Part of chromatin remodeling machinery predominantly forms H3K4me1 and H3K4me2 methylation marks at active chromatin sites where transcription and DNA repair take place (PubMed:12453419, PubMed:15960975, PubMed:19187761, PubMed:19556245, PubMed:20677832, PubMed:21220120, PubMed:25561738, PubMed:26886794). Has weak methyltransferase activity by itself, and requires other component of the MLL1/MLL complex to obtain full methyltransferase activity (PubMed:19187761, PubMed:26886794). 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' (PubMed:19187761). Binds to unmethylated CpG elements in the promoter of target genes and helps maintain them in the nonmethylated state (PubMed:20010842). Required for transcriptional activation of HOXA9 (PubMed:12453419, PubMed:20010842, PubMed:20677832). Promotes PPP1R15A-induced apoptosis (PubMed:10490642). Plays a critical role in the control of circadian gene expression and is essential for the transcriptional activation mediated by the CLOCK-BMAL1 heterodimer (By similarity). Establishes a permissive chromatin state for circadian transcription by mediating a rhythmic methylation of 'Lys-4' of histone H3 (H3K4me) and this histone modification directs the circadian acetylation at H3K9 and H3K14 allowing the recruitment of CLOCK-BMAL1 to chromatin (By similarity). Also has auto-methylation activity on Cys-3882 in absence of histone H3 substrate (PubMed:24235145).
ALL1, CXXC7, HRX, HTRX, MLL, MLL1, TRX1, KMT2A, Histone-lysine N-methyltransferase 2A, Lysine N-methyltransferase 2A, ALL-1, CXXC-type zinc finger protein 7, Cysteine methyltransferase KMT2A, Myeloid/lymphoid or mixed-lineage leukemia, Myeloid/lymphoid or mixed-lineage leukemia protein 1, Trithorax-like protein, Zinc finger protein HRX
Mouse Monoclonal KMT2A / MLL antibody. Suitable for IHC-P, Flow Cyt (Intra), WB and reacts with Human, Transfected cell lysate samples. Cited in 8 publications.
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
We can conjugate this antibody to FITC for you (please see ab150234 for details).
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The KMT2A protein also known as MLL (Mixed-Lineage Leukemia) is a large protein with a molecular weight of approximately 431 kDa. This protein functions as a histone methyltransferase which means it adds methyl groups to histone proteins specifically at the lysine 4 position on histone H3 (H3K4). This modification impacts chromatin structure and gene expression. KMT2A/MLL is chiefly expressed in hematopoietic stem cells and various tissue types indicating its broad role in regulation across the body. The protein features a SET domain a characteristic motif for proteins involved in chromatin modification.
This methyltransferase plays a significant role in regulating gene expression required for normal hematopoietic development and maintenance. KMT2A/MLL forms part of the multi-protein complex called COMPASS-like complex which is essential for its function in methylation activity. Within this complex it associates with other proteins like WDR5 RBBP5 and ASH2L that collaborate to control transcriptional elongation. KMT2A/MLL is also involved in maintaining the expression of various homeobox (HOX) genes which are critical for embryonic development and cell differentiation.
KMT2A/MLL plays an integral role in hematopoietic and developmental signaling pathways. One key pathway is the Wnt signaling pathway important for regulating stem cell pluripotency and cell fate decisions. Another pathway is the Notch signaling pathway important for cell differentiation processes. KMT2A interacts with proteins within these pathways such as CXXC1 which links it to the DNA binding properties required for regulating target gene expression.
KMT2A/MLL is closely linked to leukemia notably acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). A significant factor contributing to these associations is the gene fusions involving KMT2A often created during chromosomal translocations which result in oncogenic activation. In these leukemias it can form fusion proteins like MLL-AF4 impacting the normal regulatory activities of KMT2A. This aberrant interaction with proteins such as MEN1 can contribute to oncogenic processes by altering transcription and leading to uncontrolled cell growth.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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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) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control 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.
All lanes: Western blot - Anti-KMT2A / MLL antibody [mmN4.4] (ab32400) at 10 µg/mL
All lanes: Lysate from transfected cells overexpressing KMT2A/MLL
All lanes: Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size: 432 kDa
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.
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