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AB245738

Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free

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Rabbit Recombinant Monoclonal KMT6 / EZH2 antibody. Carrier free. Suitable for ChIC/CUT&RUN-seq, IP, Flow Cyt (Intra), WB, IHC-P and reacts with Human, Mouse samples.

View Alternative Names

KMT6, EZH2, Histone-lysine N-methyltransferase EZH2, ENX-1, Enhancer of zeste homolog 2, Lysine N-methyltransferase 6

4 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free (AB245738)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free (AB245738)

Formalin-fixed, paraffin-embedded human prostate cancer tissue stained for KMT6 / EZH2 using ab227648 at 1/100 dilution in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS#44; BSA#44; and sodium azide (ab227648)

Flow Cytometry (Intracellular) - Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free (AB245738)
  • Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free (AB245738)

Flow cytometric analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling KMT6 / EZH2 with ab227648 at 1/100 dilution (green) compared with a negative control of rabbit IgG (blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab227648).

ChIC/CUT&RUN sequencing - Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free (AB245738)
  • ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free (AB245738)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 K-562 (Human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab227648 [SP129]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab227648).

Immunoprecipitation - Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free (AB245738)
  • IP

Lab

Immunoprecipitation - Anti-KMT6 / EZH2 antibody [SP129] - BSA and Azide free (AB245738)

ab227648 (purified) at 1/20 dilution (20 μg/ml) immunoprecipitating KMT6 / EZH2 in HEK-293 whole cell lysate.
Lane 1 (input) : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab227648 & HEK-293 whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab227648 in HEK-293 whole cell lysate
For western blotting, ab227648 (purified at 1/500 dilution (0.632 μg/ml) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution were used.
Blocking and diluting buffer : 5% NFDM /TBST .

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide ab227648).

All lanes:

Immunoprecipitation - Anti-KMT6 / EZH2 antibody [SP129] (<a href='/en-us/products/primary-antibodies/kmt6-ezh2-antibody-sp129-ab227648'>ab227648</a>)

Predicted band size: 85 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

SP129

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Human

Applications

ChIC/CUT&RUN-seq, Flow Cyt (Intra), IHC-P, WB, IP

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ChICCUTRUNseq" : {"fullname" : "ChIC/CUT&RUN sequencing", "shortname":"ChIC/CUT&RUN-seq"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ChICCUTRUNseq-species-checked": "testedAndGuaranteed", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "<p></p>", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p>Primary antibody incubation for 30 minutes at 4 degrees.</p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p>Primary antibody incubation for 1 hour at room temperature.</p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p>Perform heat mediated antigen retrieval with EDTA buffer before commencing with IHC staining protocol.Primary antibody incubation for 30 minutes at room temperature.</p>" }, "Mouse": { "ChICCUTRUNseq-species-checked": "guaranteed", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "", "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p>Primary antibody incubation for 1 hour at room temperature.</p>", "IHCP-species-checked": "guaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Rat": { "ChICCUTRUNseq-species-checked": "predicted", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "", "IP-species-checked": "predicted", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Monkey": { "ChICCUTRUNseq-species-checked": "predicted", "ChICCUTRUNseq-species-dilution-info": "", "ChICCUTRUNseq-species-notes": "", "IP-species-checked": "predicted", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "predicted", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }
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Product details

ab245738 is the carrier-free version of ab227648.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A/G
Purification notes
Purified from TCS by protein A/G.
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

KMT6 also known as EZH2 (Enhancer of Zeste Homolog 2) is a protein that functions as a histone methyltransferase. It catalyzes the methylation of histone H3 on lysine 27 (H3K27) a modification associated with gene silencing. EZH2 is a component of the Polycomb Repressive Complex 2 (PRC2) which plays an important role in chromatin remodeling and transcriptional repression. The EZH2 protein has a molecular weight of approximately 87 kDa and its gene is located on chromosome 7. In terms of expression EZH2 is found in various tissues but is notably abundant in growing and dividing cells.
Biological function summary

EZH2 plays a central role in regulating gene expression during development and differentiation. It is a core component of the PRC2 complex which includes other proteins such as EED and SUZ12. Through its catalytic activity EZH2 contributes to the maintenance of the transcriptionally repressed state of genes influencing processes such as cell identity and proliferation. In many cell types EZH2-mediated gene silencing is necessary for normal development and dysregulation can lead to aberrant cellular processes.

Pathways

EZH2 participates in key regulatory networks notably the Wnt signaling pathway and the p53 pathway. Within the Wnt pathway EZH2's activity contributes to controlling gene expression that influences cellular differentiation and proliferation. Meanwhile interaction with the p53 pathway can impact the cellular response to stress and DNA damage highlighting EZH2’s regulatory dimension. Other proteins like β-catenin in the Wnt pathway interact with EZH2 reflecting its integration in these complex signaling networks.

EZH2 has been linked to cancer and other proliferative diseases. Overexpression or mutation of EZH2 is frequently observed in cancers such as prostate and breast cancer where it contributes to unchecked cell growth and tumor progression. In these contexts proteins like p53 can interact abnormally with EZH2 leading to failure in tumor suppression mechanisms. Additionally deregulated EZH2 activity is implicated in certain developmental disorders highlighting its broader impact on cellular and organismal homeostasis.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Polycomb group (PcG) protein. Catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) 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. Displays a preference for substrates with less methylation, loses activity when progressively more methyl groups are incorporated into H3K27, H3K27me0 > H3K27me1 > H3K27me2 (PubMed : 22323599, PubMed : 30923826). Compared to EZH1-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. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4 and the nuclear receptor RORA. Regulates the circadian clock via histone methylation at the promoter of the circadian genes. Essential for the CRY1/2-mediated repression of the transcriptional activation of PER1/2 by the CLOCK-BMAL1 heterodimer; involved in the di and trimethylation of 'Lys-27' of histone H3 on PER1/2 promoters which is necessary for the CRY1/2 proteins to inhibit transcription.
See full target information EZH2
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chicCutRunSequencingBooklet
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