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DNMT3A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon4 and 1 bp insertion in exon4.

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Images

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing, Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon4 and 1 bp insertion in exon4.

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Alternative names

What's included?

1 Kit
Components
Human DNMT3A knockout HeLa cell lysate
1 x 100 µg
Human wild-type HeLa cell lysate
1 x 100 µg

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DNMT3A KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon4 and 1 bp insertion in exon4.

Alternative names

Key facts

Cell type

HeLa

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon4 and 1 bp insertion in exon4.

Disease

Adenocarcinoma

Concentration
Loading...

Properties

Gene name

DNMT3A

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Storage

Shipped at conditions

Ambient - Can Ship with Ice

Appropriate long-term storage conditions

-20°C

Notes


Knockout cell lysate achieved by CRISPR/Cas9.

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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Western blot - Human DNMT3A knockout HeLa cell lysate (ab257128), expandable thumbnail

    Western blot - Human DNMT3A knockout HeLa cell lysate (ab257128)

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

    Lane 2: DNMT3A knockout HeLa cell lysate (20 μg)

    Lane 3: HEK-293 cell lysate (20 μg)

    Lane 4: Daudi cell lysate (20 μg)

    Lanes 1-4: Merged signal (red and green). Green - ab227823 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.

    ab227823 Anti-Dnmt3a antibody [EPR18455-94] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab227823 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-Dnmt3a antibody [EPR18455-94] (AB227823) at 1/1000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: Dnmt3a knockout HeLa cell lysate at 20 µg

    Lane 3: HEK-293 cell lysate at 20 µg

    Lane 4: Daudi cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (AB216773) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 102 kDa

    Observed band size: 125 kDa

    Lanes 1-4: Merged signal (red and green). Green - ab227823 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.

    ab227823 Anti-Dnmt3a antibody [EPR18455-94] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab227823 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.

  • Western blot - Human DNMT3A knockout HeLa cell lysate (ab257128), expandable thumbnail

    Western blot - Human DNMT3A knockout HeLa cell lysate (ab257128)

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

    Lane 2: DNMT3A knockout HeLa cell lysate (20 μg)

    Lane 3: HEK-293 cell lysate (20 μg)

    Lane 4: Daudi cell lysate (20 μg)

    Lanes 1-4: Merged signal (red and green). Green - ab188470 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.

    ab188470 Anti-Dnmt3a antibody [EPR18455] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab188470 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 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-Dnmt3a antibody [EPR18455] (AB188470) at 1/2000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: Dnmt3a knockout HeLa cell lysate at 20 µg

    Lane 3: HEK-293 cell lysate at 20 µg

    Lane 4: Daudi cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (AB216773) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 102 kDa

    Observed band size: 125 kDa

    Lanes 1-4: Merged signal (red and green). Green - ab188470 observed at 125 kDa. Red - loading control ab8245 observed at 37 kDa.

    ab188470 Anti-Dnmt3a antibody [EPR18455] was shown to specifically react with Dnmt3a in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261793 (knockout cell lysate ab257128) was used. Wild-type and Dnmt3a knockout samples were subjected to SDS-PAGE. ab188470 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 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.

  • Sanger Sequencing - Human DNMT3A knockout HeLa cell lysate (ab257128), expandable thumbnail

    Sanger Sequencing - Human DNMT3A knockout HeLa cell lysate (ab257128)

    Allele-1: 1 bp deletion in exon4

  • Sanger Sequencing - Human DNMT3A knockout HeLa cell lysate (ab257128), expandable thumbnail

    Sanger Sequencing - Human DNMT3A knockout HeLa cell lysate (ab257128)

    Allele-2: 1 bp insertion in exon4

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Product protocols

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

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

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