MAP2K2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
Alternative names
CFC syndrome, CFC4, Cardiofaciocutaneous syndrome, Dual specificity mitogen-activated protein kinase kinase 2, ERK activator kinase 2, FLJ26075, MAP kinase kinase 2, MAP2K2, MAPK / ERK kinase 2, MAPKK 2, MEK 2, MKK 2, MP2K2_HUMAN, Microtubule associated protein kinase kinase 2, Mitogen activated protein kinase kinase 2, Mitogen activated protein kinase kinase 2 p45, OTTHUMP00000165826, OTTHUMP00000165827, PRKMK 2
Recommended products
MAP2K2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- MAP2K2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation 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.
- Culture medium
- McCoY5a + 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
Recommended control: Human wild-type HCT116 cell line (ab288559). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
MEK2 or MAP2K2 is a protein kinase with a mass of approximately 44 kDa. This kinase part of the Mitogen-Activated Protein Kinase (MAPK) pathway primarily functions mechanically as a dual-specificity protein kinase. It phosphorylates both serine/threonine and tyrosine residues on its substrate proteins. MEK2 is largely expressed in various tissues including the brain and heart. It serves an important role in relaying extracellular signals within cells leading to diverse cellular responses.
- Biological function summary
MEK2 participates in signal transduction pathways mediating cell division differentiation and apoptosis. As a part of the MAPK signaling cascade it operates downstream of RAS and RAF proteins. MEK2 forms a complex with MEK1 another closely related kinase to ensure proper signal relay. This complex plays a significant role in the regulation of cellular responses to growth signals and stress.
- Pathways
MEK2 is intricately connected to the ERK1/2 pathway one of the key pathways it is involved in. ERK1/2 (Extracellular signal-Regulated Kinases) further transmits signals to the cell's nuclear components regulating gene expression critically. MEK2 directly phosphorylates and activates ERK1 and ERK2 facilitating their role in transcription cell cycle regulation and maintenance of cell integrity. The interaction of MEK2 with proteins like BRAF and CRAF along the MAPK pathway amplifies the signal transduction for various growth factors and hormones.
- Associated diseases and disorders
MEK2 has a notable association with cancer particularly melanoma and colorectal cancer. It often becomes aberrantly activated leading to unregulated cell proliferation. The connection of MEK2 to ERK1/2 proteins in these malignancies makes it a potential therapeutic target. Furthermore MEK2 mutations can contribute to Noonan syndrome a disorder that affects multiple body systems marking its relationship with genetic abnormalities and developmental issues. Researchers have been investigating MEK inhibitors to target these disorders highlighting the importance of understanding MEK2's role in disease contexts.
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2 product images
Western blot - Human MAP2K2 knockout HCT116 cell line (ab286599)
Western blot: Anti-MAP2K2 antibody [Y78] (Anti-MEK2 antibody [Y78] ab32517) staining at 1/10000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-MEK2 antibody [Y78] ab32517 was shown to bind specifically to MAP2K2. A band was observed at 44 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in MAP2K2 knockout cell line. To generate this image, wild-type and MAP2K2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-MEK2 antibody [Y78] (Anti-MEK2 antibody [Y78] ab32517) at 1/10000 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human MAP2K2 knockout HCT116 cell line (ab286599)
Lane 2: MAP2K2 knockout HCT 116 cell lysate at 20 µg
Lane 3: Wild-type HEK-293T ab255553 cell lysate at 20 µg
Lane 4: MAP2K2 knockout HEK-293T ab261003 cell lysate at 20 µg
SecondaryLanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 44 kDa
Next Generation Sequencing - Human MAP2K2 knockout HCT116 cell line (ab286599)
92 bp deletion in exon 7, CCDS12120.1
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