Human MAP3K11 (MLK3) knockout A549 cell line
- Advanced Validation
- What is this?
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MAP3K11 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 5 and 2 bp deletion in exon 5 and 7 bp deletion in exon 5. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
2610017K16Rik, EC 2.7.11.25, M3K11_HUMAN, MEKK11, MGC17114, Map3k11, Mitogen-activated protein kinase kinase kinase 11, Mixed lineage kinase 3, Mixed lineage protein kinase 3, PTK1, Protein tyrosine kinase PTK1, RHOE, SH3 domain containing proline rich kinase, SPRK, Src-homology 3 domain-containing proline-rich kinase
- WB
Lab
Western blot - Human MAP3K11 (MLK3) knockout A549 cell line (AB267168)
Lanes 1- 2 : Merged signal (red and green). Green - ab51068 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab51068 was shown to 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 A549 and MAP3K11 knockout A549 cell lysates 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) overnight at 4°C at a 1 in 5000 dilution and a 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 (ab267168)
Predicted band size: 93 kDa
Observed band size: 105 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell line (AB267168)
Allele-1 : 7 bp deletion in exon5
- Sanger seq
Unknown
Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell line (AB267168)
Allele-3 : 1 bp insertion in exon 5.
- Sanger seq
Unknown
Sanger Sequencing - Human MAP3K11 (MLK3) knockout A549 cell line (AB267168)
Allele-2 : 2 bp deletion in exon 5.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
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
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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