AB257080

Human EHMT2 (G9A) knockout HeLa cell lysate

Name: Human EHMT2 (G9A) knockout HeLa cell lysate (ab257080) | Abcam
Brand: Abcam Limited
SKU: AB257080

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EHMT2 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon7 and 370 bp deletion in exon7.

View Alternative Names

Lysine N-methyltransferase 1C, KMT1C, G9A, BAT8, H3-K9-HMTase 3, NG36, Euchromatic histone-lysine N-methyltransferase 2, EHMT2, Histone H3-K9 methyltransferase 3, C6orf30, Histone-lysine N-methyltransferase EHMT2, Protein G9a, HLA-B-associated transcript 8

4 Images

Lab

Western blot - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : EHMT2 knockout HeLa cell lysate (20 μg)

Lane 3 : HEK-293 cell lysate (20 μg)

Lane 4 : HepG2 cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab133482 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.

ab133482 Anti-EHMT2/G9A antibody [EPR4019(2)] was shown to specifically react with EHMT2/G9A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265149 (knockout cell lysate ab257080) was used. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab133482 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-EHMT2/G9A antibody [EPR4019(2)] - ChIP Grade (<a href='/en-us/products/primary-antibodies/ehmt2-g9a-antibody-epr40192-chip-grade-ab133482'>ab133482</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

EHMT2/G9A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human EHMT2 (G9A) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ehmt2-g9a-knockout-hela-cell-line-ab265149'>ab265149</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 132 kDa

Observed band size: 160 kDa

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Lab

Western blot - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : EHMT2 knockout HeLa cell lysate (20 μg)

Lane 3 : HEK-293 cell lysate (20 μg)

Lane 4 : HepG2 cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab185050 observed at 160 kDa. Red - loading control ab8245 observed at 37 kDa.

ab185050 Anti-EHMT2/G9A antibody [EPR18894] was shown to specifically react with EHMT2/G9A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265149 (knockout cell lysate ab257080) was used. Wild-type and EHMT2/G9A knockout samples were subjected to SDS-PAGE. ab185050 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-EHMT2/G9A antibody [EPR18894] - ChIP Grade (<a href='/en-us/products/primary-antibodies/ehmt2-g9a-antibody-epr18894-chip-grade-ab185050'>ab185050</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

EHMT2/G9A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human EHMT2 (G9A) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ehmt2-g9a-knockout-hela-cell-line-ab265149'>ab265149</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Predicted band size: 132 kDa

Observed band size: 160 kDa

false

Sanger Sequencing - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Sanger seq

Unknown

Sanger Sequencing - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Allele-2 : 370 bp deletion in exon7

Sanger seq

Unknown

Sanger Sequencing - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Allele-1 : 2 bp deletion in exon7

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon7 and 370 bp deletion in exon7.

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name

EHMT2

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing

Shipped at conditions

Ambient - Can Ship with Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

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Supplementary information

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

EHMT2 also known as G9a is a methyltransferase enzyme with a molecular mass of approximately 165 kDa. It modifies histones by adding methyl groups to lysine 9 on histone H3 which results in chromatin structure changes and influences gene expression. EHMT2/G9a expresses in various tissues including brain heart and liver showing its widespread role in different physiological processes. As a well-studied protein researchers often analyze EHMT2 activity using assays like ELISA and western blot.

Biological function summary

EHMT2/G9a regulates gene expression by altering chromatin accessibility. It is part of a chromatin remodeling complex interacting with proteins such as ANMTA02. Through its methyltransferase activity it maintains an epigenetic silence state important for embryonic development differentiation and cellular identity. EHMT2/G9a plays a significant role in the repression of transcriptional activity of tumor suppressor genes and developmental genes.

Pathways

G9a functions in epigenetic signaling pathways and influences processes like the cell cycle and DNA damage repair. It interacts with proteins in the Polycomb repressive complex such as EZH2 to regulate gene silencing. Additionally in the DNA damage response pathway G9a can associate with histone deacetylases to stabilize chromatin and prevent unwarranted gene expression.

Aberrant regulation of EHMT2/G9a associates with cancer and neurodevelopmental disorders. In various cancers such as leukemia G9a suppresses tumor suppressor genes aiding tumor progression. Connections between G9a and proteins like DNMT1 highlight its role in maintaining a hypermethylation state in cancerous cells. In neurodevelopmental disorders altered expression of G9a affects synaptic plasticity linking it with cognitive impairments and autism spectrum disorders.

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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Human EHMT2 (G9A) knockout HeLa cell lysate

Western blot - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Western blot - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Sanger Sequencing - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Sanger Sequencing - Human EHMT2 (G9A) knockout HeLa cell lysate (AB257080)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

Complementary Products

Primary Antibodies

Proteins & Peptides

Proteins & Peptides

Primary Antibodies

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Product promise