MAP3K2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 7 and 2 bp deletion in exon 7.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 7 and 2 bp deletion in exon 7
Alternative names
M3K2_HUMAN, MAPK/ERK kinase kinase 2, MEK kinase 2, MEKK2b, Map3k2, Mitogen-activated protein kinase kinase kinase 2
Recommended products
MAP3K2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 7 and 2 bp deletion in exon 7.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 7 and 2 bp deletion in exon 7
- Antibiotic resistance
- Puromycin 1µg/mL
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- MAP3K2
- 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: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
MEKK2 also known as MAP3K2 is a kinase enzyme with an approximate mass of 70 kDa. It functions as part of the MAP kinase signaling pathway where it phosphorylates downstream proteins playing an essential role in transmitting signals within cells. This protein is mainly expressed in tissues such as the brain heart and liver. It acts by activating specific MAP kinase pathways facilitating various cellular responses to external stimuli.
- Biological function summary
This kinase serves as a critical mediator in cellular processes like proliferation differentiation and apoptosis. MEKK2 forms part of larger signaling complexes including the MAP3K complex where it regulates multiple cellular activities. Through its kinase activity MEKK2 passes on signals that affect gene expression and cellular stress responses impacting how cells react to their environment.
- Pathways
MEKK2 functions prominently in the MAPK signaling and NF-kB pathways. These pathways involve signal transduction mechanisms that MEKK2 influences through its interactions with related proteins such as MEK5 and ERK5. By modulating these pathways MEKK2 participates in a variety of cellular responses including inflammation and immune responses.
- Associated diseases and disorders
MEKK2 has associations with conditions like cancer and inflammatory diseases. Aberrant expression of MEKK2 can affect the behavior of cancer cells due to its influence on the MAPK pathway's signaling leading to unchecked cell growth. Additionally MEKK2 interacts with proteins like NF-kB which are important in inflammation thereby implicating it in disorders related to immune system dysfunctions.
Product promise
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3 product images
Western blot - Human MAP3K2 (MEKK2) knockout A549 cell line (ab267152)
Lanes 1- 2: Merged signal (red and green). Green - Anti-MEKK2 antibody [EP626Y] ab33918 observed at 75 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-MEKK2 antibody [EP626Y] ab33918 was shown to react with MEKK2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267152 (knockout cell lysate Human MAP3K2 (MEKK2) knockout A549 cell lysate ab257521) was used. Wild-type A549 and MAP3K2 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. Anti-MEKK2 antibody [EP626Y] ab33918 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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-MEKK2 antibody [EP626Y] (Anti-MEKK2 antibody [EP626Y] ab33918) at 1/10000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: MAP3K2 knockout A549 cell lysate at 20 µg
Lane 2: Western blot - Human MAP3K2 (MEKK2) knockout A549 cell line (ab267152)
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 75 kDa
Sanger Sequencing - Human MAP3K2 (MEKK2) knockout A549 cell line (ab267152)
Allele-2: 1 bp insertion in exon 7.
Sanger Sequencing - Human MAP3K2 (MEKK2) knockout A549 cell line (ab267152)
Allele-1: 2 bp deletion in exon7
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