Human MAP2K2 knockout HCT116 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
MAP2K2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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
- WB
Lab
Western blot - Human MAP2K2 knockout HCT116 cell line (AB286599)
Western blot : Anti-MAP2K2 antibody [Y78] (ab32517) staining at 1/10000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/mek2-antibody-y78-ab32517'>ab32517</a>) 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
Secondary
Lanes 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
Observed band size: 44 kDa
false
- NGS
Lab
Next Generation Sequencing - Human MAP2K2 knockout HCT116 cell line (AB286599)
92 bp deletion in exon 7, CCDS12120.1
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 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.
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.
Culture medium
McCoY5a + 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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Complementary Products
Primary Antibodies
AB32517
20ul selling size|KO Validated|RabMAb|Recombinant
![Flow Cytometry (Intracellular) - Anti-MEK2 antibody [Y78] (AB32517)](https://content.abcam.com/products/images/mek2-antibody-y78-ab32517--flow-cytometry-intracellular-img71584.jpg)
- 4
- 3
Primary Antibodies
AB76125
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JNK2 antibody [EP1595Y] (AB76125)](https://content.abcam.com/products/images/jnk2-antibody-ep1595y-ab76125--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img139191.gif)
- 5
- 1
![Flow Cytometry (Intracellular) - Anti-hHR23A antibody [EPR4818] (AB108592)](https://content.abcam.com/products/images/hhr23a-antibody-epr4818-ab108592--flow-cytometry-intracellular-img50725.jpg)
Primary Antibodies
AB205926
Anti-p38 gamma/MAPK12 antibody [EPR6528(N)]
RabMAb|Recombinant|KO Validated|Trial Size
![Immunocytochemistry/ Immunofluorescence - Anti-p38 gamma/MAPK12 antibody [EPR6528(N)] (AB205926)](https://content.abcam.com/products/images/p38-gamma-mapk12-antibody-epr6528n-ab205926--immunocytochemistry-immunofluorescence-img69322.jpg)
- 0
- 0
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com