MAP4K3 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
Images
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1
Alternative names
Germinal center kinase like kinase, Germinal center kinase-related protein kinase, MAP4K3, MAPK/ERK kinase kinase kinase 3, MAPKKKK3, MEK kinase kinase 3, MEKKK 3, RAB8IPL1, mitogen-activated protein kinase kinase kinase kinase 3
Recommended products
MAP4K3 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Form
Liquid
- Knockout validation
Sanger Sequencing
- Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
MAP4K3
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing
- Zygosity
Homozygous
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 2 US: 2
- Adherent/suspension
Adherent
- Gender
Female
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
DMEM (High Glucose) + 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 long-term storage conditions
-196°C
Notes
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255448). 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.
Product promise
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Full details and terms and conditions can be found here:
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2 product images
Western blot - Human MAP4K3 (GLK) knockout HeLa cell line (ab264796)
Lane 1: Wild-type HeLa cell lysate (20ug)
Lane 2: MAP4K3 knockout HeLa cell lysate(20ug)
Lane 3: HepG2 cell lysate (20ug)
Lane 4: Daudi cell lysate (20ug)
Lanes 1-2: Merged signal (red and green). Green - MAP4K3 (D1L4G) Rabbit monoclonal antibody observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
MAP4K3 (D1L4G) Rabbit monoclonal antibody was shown to specifically react with MAP4K3 in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab264796 (knockout cell lysate Human MAP4K3 (GLK) knockout HeLa cell lysate ab258956) lane below 100kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and MAP4K3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. MAP4K3 (D1L4G) Rabbit monoclonal antibody and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4oC at 1 in 1000 and 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: MAP4K3 (D1L4G) Rabbit mAb at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: MAP4K3 knockout HeLa cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
Performed under reducing conditions.
Sanger Sequencing - Human MAP4K3 (GLK) knockout HeLa cell line (ab264796)
Homozygous: Insertion of the selection cassette in exon 1.
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