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AB257518

Human MAP3K11 (MLK3) knockout A549 cell lysate

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MAP3K11 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 insertion in exon5 and 2 bp deletion in exon5 and 7 bp deletion in exon5.
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Western blot - Human MAP3K11 (MLK3) knockout A549 cell lysate (AB257518)
  • WB

Lab

Western blot - Human MAP3K11 (MLK3) knockout A549 cell lysate (AB257518)

Lane 1 : Wild-type A549 cell lysate (20µg)

Lane 2 : MAP3K11 knockout A549 cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab51068 observed at 105 kDa. Red - loading control ab8245 observed at 37 kDa.

ab51068 Anti-MLK3 antibody [EP1460Y] was shown to specifically react with MLK3 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267168 (knockout cell lysate ab257518) was used. Wild-type and MLK3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51068 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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-MLK3 antibody [EP1460Y] (<a href='/en-us/products/primary-antibodies/mlk3-antibody-ep1460y-ab51068'>ab51068</a>) at 1/5000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

MAP3K11 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human MAP3K11 (MLK3) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-map3k11-mlk3-knockout-a549-cell-line-ab267168'>ab267168</a>)

Predicted band size: 93 kDa

Observed band size: 105 kDa

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Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell lysate (AB257518)
  • Sanger seq

Unknown

Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell lysate (AB257518)

Allele-1 : 7 bp deletion in exon5

Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell lysate (AB257518)
  • Sanger seq

Unknown

Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell lysate (AB257518)

Allele-3 : 1 bp insertion in exon5

Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell lysate (AB257518)
  • Sanger seq

Unknown

Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell lysate (AB257518)

Allele-2 : 2 bp deletion in exon5

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon5 and 2 bp deletion in exon5 and 7 bp deletion in exon5.

Disease

Carcinoma

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Complementary Products

Primary Antibodies

AB51068

Anti-MLK3 antibody [EP1460Y]

RabMAb|Recombinant|KO Validated|Trial Size|Trial Size

Immunocytochemistry/ Immunofluorescence - Anti-MLK3 antibody [EP1460Y] (AB51068)

  • 0
  • 0
Proteins & Peptides

AB125630

Recombinant human MLK3 protein

Functional Studies - Recombinant human MLK3 protein (AB125630)

  • 0
  • 0
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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, 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.

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What's included?

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Properties and storage information

Gene name
MAP3K11
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.

MLK3 also known as MEKK11 is a kinase widely expressed in human tissues. This protein with a molecular mass of approximately 97 kDa functions as a mitogen-activated protein kinase kinase kinase (MAP3K). MLK3 mechanically activates the c-Jun N-terminal kinase (JNK) and p38 MAP kinase pathways by phosphorylating downstream MAP2Ks. It is involved in various cell processes including apoptosis and inflammation.
Biological function summary

MLK3 plays a critical role in cellular response to stress and inflammation. It acts as part of a signaling complex that regulates the JNK and p38 pathways key mediators in immune responses. These pathways control the production of pro-inflammatory cytokines and the cellular stress response. Through its kinase activity MLK3 supports the activation of transcription factors such as AP-1 which drive gene expression changes needed for its biological functions.

Pathways

MLK3 is intricately involved in the MAPK signaling cascade. It connects with other MAP3Ks like MLK1 and MLK2 contributing to the complexity of MAPK signaling. MLK3’s participation in the JNK and p38 pathways links it to cytokine signaling and cellular stress responses. These pathways are significant for maintaining cellular homeostasis and coordinating responses to external stimuli.

MLK3 has implications in cancer and neurodegenerative diseases. In cancer MLK3 modulates pathways that influence cell proliferation and survival potentially leading to tumor growth and metastasis when dysregulated. Similarly altered MLK3 signaling is observed in neurodegenerative diseases possibly due to its role in managing cellular stress. MLK3 interacts with proteins like JNK and p38 which are important in disease mechanisms and progression.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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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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