Human MAP3K2 (MEKK2) knockout A549 cell line
- Advanced Validation
- What is this?
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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 4 bp deletion in exon 7.
View Alternative Names
M3K2_HUMAN, MAPK/ERK kinase kinase 2, MEK kinase 2, MEKK2b, Map3k2, Mitogen-activated protein kinase kinase kinase 2
- WB
Lab
Western blot - Human MAP3K2 (MEKK2) knockout A549 cell line (AB267153)
Lanes 1- 2 : Merged signal (red and green). Green - ab33918 observed at 75 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab33918 was shown to react with MEKK2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267153 (knockout cell lysate ab257522) 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. ab33918 and 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 (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-MEKK2 antibody [EP626Y] (<a href='/en-us/products/primary-antibodies/mekk2-antibody-ep626y-ab33918'>ab33918</a>) 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 (ab267153)
Predicted band size: 70 kDa
Observed band size: 75 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MAP3K2 (MEKK2) knockout A549 cell line (AB267153)
Allele-2 : 1 bp insertion in exon 7.
- Sanger seq
Unknown
Sanger Sequencing - Human MAP3K2 (MEKK2) knockout A549 cell line (AB267153)
Allele-1 : 4 bp deletion in exon7
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab255450). 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, 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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
![Western blot - Anti-MEKK2 antibody [EP626Y] (AB33918)](https://content.abcam.com/products/images/mekk2-antibody-ep626y-ab33918--western-blot-img100812.jpg)
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