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AB258489

Human L3MBTL3 knockout HEK-293T cell lysate

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L3MBTL3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 28 bp deletion in exon9.
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Western blot - Human L3MBTL3 knockout HEK-293T cell lysate (AB258489)
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Western blot - Human L3MBTL3 knockout HEK-293T cell lysate (AB258489)

Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2 : L3MBTL3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate (20 ug)

ab175232 was shown to specifically react with L3MBTL3 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266172 (knockout cell lysate ab258489) was used. Wild-type and L3MBTL3 knockout samples were subjected to SDS-PAGE. ab175232 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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-L3MBTL3 antibody [EPR11854] (<a href='/en-us/products/primary-antibodies/l3mbtl3-antibody-epr11854-ab175232'>ab175232</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

L3MBTL3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human L3MBTL3 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-l3mbtl3-knockout-hek-293t-cell-line-ab266172'>ab266172</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HepG2 (Human liver hepatocellular carcinoma cell line) whole 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 1/10000 dilution

Predicted band size: 88 kDa

Observed band size: 88 kDa

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Sanger Sequencing - Human L3MBTL3 knockout HEK-293T cell lysate (AB258489)
  • Sanger seq

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Sanger Sequencing - Human L3MBTL3 knockout HEK-293T cell lysate (AB258489)

Homozygous : 28 bp deletion in exon9

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 28 bp deletion in exon9.

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Complementary Products

Primary Antibodies

AB249882

Anti-L3MBTL3 antibody [EPR11854] - BSA and Azide free

RabMAb|Recombinant|KO Validated

Western blot - Anti-L3MBTL3 antibody [EPR11854] - BSA and Azide free (AB249882)

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Cell Lines & Lysates

AB266172

Human L3MBTL3 knockout HEK-293T cell line

Advanced Validation

Western blot - Human L3MBTL3 knockout HEK-293T cell line (AB266172)

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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
L3MBTL3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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.

L3MBTL3 also known as Lethal(3) Malignant Brain Tumor-Like Protein 3 is a member of the polycomb group proteins which regulate gene expression by remodeling chromatin. It has a molecular weight of approximately 69 kDa. L3MBTL3 is widely expressed in various tissues with significant levels in the brain and hematopoietic cells. This protein plays a role in chromatin compaction through its ability to recognize and bind mono- and dimethylated lysine residues on histones. These activity patterns classify it as a histone reader influencing the accessibility of transcriptionally active regions.
Biological function summary

L3MBTL3 affects cellular processes by managing chromatin structure which in turn regulates gene expression. It participates in chromatin remodeling as part of large multi-protein complexes. By interacting with different components L3MBTL3 modulates the transcription of oncogenes and tumor suppressors. It contributes to important cellular processes including cell-cycle progression and differentiation by establishing particular transcriptional profiles. The protein's role in these processes highlights its importance in maintaining normal cell function.

Pathways

L3MBTL3 participates in pathways that involve chromatin modification and gene silencing. It is associated with the Polycomb Repressive Complex 2 (PRC2) pathway which plays a role in maintaining transcriptional repression. This pathway involves key proteins such as EZH2 and SUZ12 which work alongside L3MBTL3 to modify histone marks thereby silencing target genes. This positioning within pathways makes it an integral component of transcriptional regulation by ensuring genes are restricted as necessary for proper development and cellular activity.

L3MBTL3 has associations with cancer and hematological abnormalities. Its dysregulation has links to certain types of leukemia where abnormal expression can lead to uncontrolled cell proliferation. Another condition tied to L3MBTL3 is brain malignancies due to perturbations in its normal function within neuronal cells. Interactions with proteins like EZH2 further connect L3MBTL3 to cancer pathways emphasizing the relevance of its careful regulation in maintaining cellular health and preventing disease.
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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

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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'.

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