Recombinant Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal (ab191080)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR9307(2)] to KMT6 / EZH2 - N-terminal
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal
See all KMT6 / EZH2 primary antibodies -
Description
Rabbit monoclonal [EPR9307(2)] to KMT6 / EZH2 - N-terminal -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-Pmore details
Unsuitable for: ChIP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Human breast carcinoma tissue; HeLa cells. WB: HEK293 cell lysate, Ms testis tissue lysate, mouse and rat spleen tissue lysate
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR9307(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab191080 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 |
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Flow Cyt (Intra) |
1/90.
Purified format. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/500. Detects a band of approximately 93 kDa (predicted molecular weight: 85 kDa).
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ICC/IF | (1) |
1/250.
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IHC-P |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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Flow Cyt (Intra)
1/90. Purified format. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/500. Detects a band of approximately 93 kDa (predicted molecular weight: 85 kDa). |
ICC/IF
1/250. |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Polycomb group (PcG) protein. Catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' and 'Lys-27' of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. -
Tissue specificity
Expressed in many tissues. Overexpressed in numerous tumor types including carcinomas of the breast, colon, larynx, lymphoma and testis. -
Sequence similarities
Belongs to the histone-lysine methyltransferase family. EZ subfamily.
Contains 1 SET domain. -
Developmental stage
Expression decreases during senescence of embryonic fibroblasts (HEFs). Expression peaks at the G1/S phase boundary. -
Post-translational
modificationsPhosphorylated by AKT1. Phosphorylation by AKT1 reduces methyltransferase activity. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 2146 Human
- Entrez Gene: 14056 Mouse
- Entrez Gene: 312299 Rat
- Omim: 601573 Human
- SwissProt: Q15910 Human
- SwissProt: Q61188 Mouse
- Unigene: 444082 Human
- Unigene: 246688 Mouse
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Alternative names
- Enhancer of zeste 2 antibody
- enhancer of zeste 2 polycomb repressive complex 2 subunit antibody
- Enhancer of zeste homolog 2 (Drosophila) antibody
see all
Images
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All lanes : Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal (ab191080) at 1/500 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : ezh2 CRISPR-Cas9 edited MCF7 cell lysate
Lane 3 : HEK-293 cell lysate
Lane 4 : Mouse Testis cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-KMT6 / EZH2 antibody [EPR9307(2)] - N-terminal staining at 1/500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab191080 was shown to bind specifically to KMT6 / EZH2. A band was observed at 90 kDa (green arrow) in wild-type MCF7 cell lysates with no signal observed at this size in ezh2 CRISPR-Cas9 edited cell line ab281611 (CRISPR-Cas9 edited cell lysate ab282963). The band observed in the CRISPR-Cas9 edited lysate lane below 90 kDa (yellow arrow) is likely to represent a truncated form of KMT6 / EZH2. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and ezh2 CRISPR-Cas9 edited MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: KMT6 / EZH2 knockout HAP1 cell lysate (20 µg)
Lane 3: HEK293 cell lysate (20 µg)
Lane 4: Ms testis tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab191080 observed at 93 kDa. Red - loading control, ab8245, observed at 37 kDa.ab191080 was shown to recognize KMT6 / EZH2 when KMT6 / EZH2 knockout samples were used, along with additional cross-reactive bands. Wild-type and KMT6 / EZH2 knockout samples were subjected to SDS-PAGE. ab191080 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling KMT6 / EZH2 with ab191080 at 1/250 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin (inset: negative control).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of HeLa cells (4% Paraformaldehyde-fixed; 0.1% tritonX-100-permeabilized) labeling KMT6 / EZH2 with ab191080 at 1/250 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) secondary at 1/200 dilution and counter-stained with DAPI (blue).
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Lane 1: HEK293 cell lysate (20 μg)
Lane 2: Mouse spleen tissue lysate (20 μg)
Lane 3: Rat spleen tissue lysate (20 μg)
Lanes 1 - 3: Merged signal (red and green). Green - ab191080 observed at 93 kDa. Red - loading control, ab8245, observed at 37 kDa.
Human, mouse and rat extracts were subjected to SDS-PAGE. ab191080 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
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ab191080 staining KMT6 / EZH2in the human cell line Jurkat (human acute T cell leukemia) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/90. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (38)
ab191080 has been referenced in 38 publications.
- Niwa Y et al. Cyclin D1 Binding Protein 1 Responds to DNA Damage through the ATM-CHK2 Pathway. J Clin Med 11:N/A (2022). PubMed: 35160302
- Zhao T et al. Folic Acid Attenuates Glial Activation in Neonatal Mice and Improves Adult Mood Disorders Through Epigenetic Regulation. Front Pharmacol 13:818423 (2022). PubMed: 35197855
- Wang L et al. Combined Detection of RUNX3 and EZH2 in Evaluating Efficacy of Neoadjuvant Therapy and Prognostic Value of Middle and Low Locally Advanced Rectal Cancer. Front Oncol 12:713335 (2022). PubMed: 35280723
- Guo SM et al. Regulatory roles of alternative splicing at Ezh2 gene in mouse oocytes. Reprod Biol Endocrinol 20:99 (2022). PubMed: 35791029
- Ye J et al. Long non-coding RNA FOXP4-AS1 facilitates the biological functions of hepatocellular carcinoma cells via downregulating ZC3H12D by mediating H3K27me3 through recruitment of EZH2. Cell Biol Toxicol 38:1047-1062 (2022). PubMed: 34545456