MAP3K7 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 6.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 6
Alternative names
M3K7_HUMAN, MAP3K 7, MEKK7, Mitogen-activated protein kinase kinase kinase 7, TAK1, TGF-beta-activated kinase 1, TGF1a, Transforming growth factor-beta-activated kinase 1
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MAP3K7 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 6.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 6
- Concentration
Properties
- Gene name
- MAP3K7
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
- 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 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
TAK1 also known as MAP3K7 (Mitogen-activated protein kinase kinase kinase 7) is an important protein kinase that weighs approximately 63 kDa. This protein is expressed in various tissues and cells including HEK 293T cells. TAK1 plays a mechanical role as a part of the MAPK signaling cascade. It phosphorylates and activates downstream kinases which is key to transmitting cellular signals that regulate responses to external stimuli like cytokines and stress.
- Biological function summary
TAK1 is involved in several cellular processes necessary for maintaining balance and responding to stress. TAK1 forms a complex with proteins like TAB1 TAB2 and TAB3 which are important for its activation and signaling function. This complex facilitates TAK1's involvement in inflammation and immune response indicating its significance in mediating cellular survival and apoptosis.
- Pathways
TAK1 operates within the NF-κB and MAPK pathways two critical routes for cellular response to inflammation and stress. In the NF-κB pathway TAK1 activates IKK which triggers the degradation of IκB freeing NF-κB to move into the nucleus and activate transcription. In the MAPK pathway TAK1 directly influences JNK and p38 cascades revealing regulatory connections to proteins like MEK and ERK.
- Associated diseases and disorders
TAK1 has been linked to conditions such as cancer and inflammatory diseases. In cancers aberrant TAK1 activity can lead to enhanced cell proliferation and survival. Moreover inflammation-related disorders can arise from malfunctioning TAK1 signaling demonstrating its connection to proteins like TNF receptor-associated factors which are involved in inflammatory responses. Understanding TAK1's role in these disorders may pave the way for developing potential therapeutic interventions.
Product promise
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Full details and terms and conditions can be found here:
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3 product images
Cell Culture - Human MAP3K7 (TAK1) knockout HEK-293T cell line (ab266555)
Representative images MAP3K7 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
Western blot - Human MAP3K7 (TAK1) knockout HEK-293T cell line (ab266555)
Lanes 1- 2: Merged signal (red and green). Green - Anti-TAK1 antibody [EPR5984] ab109526 observed at 72 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.Anti-TAK1 antibody [EPR5984] ab109526 was shown to react with TAK1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266555 (knockout cell lysate Human MAP3K7 (TAK1) knockout HEK-293T cell lysate ab256984) was used. Wild-type HEK-293T and MAP3K7 knockout HEK-293T 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-TAK1 antibody [EPR5984] ab109526 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4° at a 1 in 1000 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-TAK1 antibody [EPR5984] (Anti-TAK1 antibody [EPR5984] ab109526) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: MAP3K7 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human MAP3K7 (TAK1) knockout HEK-293T cell line (ab266555)
Performed under reducing conditions.
Predicted band size: 35 kDa, 36 kDa, 43 kDa, 47 kDa, 60 kDa, 67 kDa, 74 kDa
Observed band size: 36 kDa, 60 kDa, 72 kDa, 95 kDa
Sanger Sequencing - Human MAP3K7 (TAK1) knockout HEK-293T cell line (ab266555)
Homozygous: 2 bp insertion in exon 6
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