Recombinant Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
![ChIP - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)](https://www.abcam.com/ps/products/191/ab191250/Images/ab191250-6-AntiKMT6-EZH2-antibody-EPR20108--ChIP-GradeChIP.jpg "ChIP - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20108] to KMT6 / EZH2 - ChIP Grade
- Suitable for: ChIP, WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade
See all KMT6 / EZH2 primary antibodies -
Description
Rabbit monoclonal [EPR20108] to KMT6 / EZH2 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: ChIP, WB, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MCF7, HeLa, U-2 OS and HEK-293 whole cell lysates; Wild-type HAP1 cell lysate. IP: HeLa whole cell lysate. ChIP: Chromatin from MDA-MB-231 cells.
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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: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20108 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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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 ab191250 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 |
|---|---|---|
| ChIP | (2) |
Use 5 µg for 25 µg of chromatin.
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| WB | (2) |
1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 85 kDa).
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| IP |
1/30.
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| Notes |
|---|
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ChIP
Use 5 µg for 25 µg of chromatin. |
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WB
1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 85 kDa). |
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IP
1/30. |
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
- Omim: 601573 Human
- SwissProt: Q15910 Human
- Unigene: 444082 Human
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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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ChIP - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
Chromatin was prepared from MDA-MB-231 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab191250 (red), and 20µl of Anti rabbit IgG sepharose beads. 5μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Cell line and primers selection based on literature (PMID: 21880597).
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![Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)](https://www.abcam.com/ps/products/191/ab191250/Images/ab191250-276190-anti-kmt6-ezh2-antibody-epr20108-chip-grade-western-blot.jpg)
Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)All lanes : Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : KMT6 / EZH2 knockout HAP1 cell lysate
Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 85 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Merged signal (red and green). Green - ab191250 observed at 93 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab191250 was shown to specifically recognize KMT6/EZH2 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when KMT6/EZH2 knockout samples were examined. Wild-type and KMT6/EZH2 knockout samples were subjected to SDS-PAGE. ab191250 and ab8245 (loading control to GAPDH) were diluted at 1/1000 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 hour at room temperature before imaging.
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![Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)](https://www.abcam.com/ps/products/191/ab191250/Images/ab191250-276188-anti-kmt6-ezh2-antibody-epr20108-chip-grade-western-blot.jpg)
Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)All lanes : Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250) at 1/5000 dilution
Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 85 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)](https://www.abcam.com/ps/products/191/ab191250/Images/ab191250-276187-anti-kmt6-ezh2-antibody-epr20108-chip-grade-western-blot.jpg)
Western blot - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250) at 1/1000 dilution + U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 85 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Immunoprecipitation - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)](https://www.abcam.com/ps/products/191/ab191250/Images/ab191250-276186-anti-kmt6-ezh2-antibody-epr20108-chip-grade-immunoprecipitation.jpg)
Immunoprecipitation - Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)KMT6 / EZH2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab191250 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab191250 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (Input).
Lane 2: ab191250 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191250 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)](https://www.abcam.com/ps/products/191/ab191250/Images/ab191250-7-benefits-of-recombinant-antibodies.png)
Anti-KMT6 / EZH2 antibody [EPR20108] - ChIP Grade (ab191250)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (27)
ab191250 has been referenced in 27 publications.
- Li R et al. LINC01116 Promotes Doxorubicin Resistance in Osteosarcoma by Epigenetically Silencing miR-424-5p and Inducing Epithelial-Mesenchymal Transition. Front Pharmacol 12:632206 (2021). PubMed: 33762953
- Kuang Y et al. Inhibition of microRNA let-7b expression by KDM2B promotes cancer progression by targeting EZH2 in ovarian cancer. Cancer Sci 112:231-242 (2021). PubMed: 33091189
- Qu S et al. EZH2 is involved in psoriasis progression by impairing miR-125a-5p inhibition of SFMBT1 and leading to inhibition of the TGFß/SMAD pathway. Ther Adv Chronic Dis 12:2040622320987348 (2021). PubMed: 33948156
- Yang L et al. PRMT5 functionally associates with EZH2 to promote colorectal cancer progression through epigenetically repressing CDKN2B expression. Theranostics 11:3742-3759 (2021). PubMed: 33664859
- Liu Z et al. EZH2-mediated epigenetic suppression of SH3BGRL potently inhibits lung cancer progression. Biochem Biophys Res Commun 548:53-59 (2021). PubMed: 33631674
