MAPK8 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Pro59 of the WT protein
Frameshift: 100%.
Images
Key facts
- Cell type
- U-2 OS
- Species or organism
- Human
- Tissue
- Bone
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Pro59 of the WT protein Frameshift: 100%
Alternative names
AI849689, C-JUN kinase 1, EC 2.7.11.24, JNK, JNK 1, JNK-46, JNK1A2, JNK21B1/2, MAP kinase 8, MAPK 8, MK08_HUMAN, Mitogen-activated protein kinase 8, PRKM 8, Protein kinase JNK1, Protein kinase, mitogen-activated, 8, SAPK 1, SAPK gamma, Stress-activated protein kinase 1, Stress-activated protein kinase JNK1, c-Jun N-terminal kinase 1, p54 gamma
Recommended products
MAPK8 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Pro59 of the WT protein
Frameshift: 100%.
Key facts
- Cell type
- U-2 OS
- Species or organism
- Human
- Tissue
- Bone
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Mutation description
- Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Pro59 of the WT protein Frameshift: 100%
- Disease
- Osteosarcoma
- Concentration
Properties
- Gene name
- MAPK8
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- 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
- 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 duration
- 1-2 weeks
- 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
JNK1 also known as c-Jun N-terminal kinase 1 is a member of the mitogen-activated protein kinase (MAPK) family with a molecular weight of approximately 46 kDa. It is expressed in various tissues throughout the body including the brain heart liver and skeletal muscle. JNK1 exists in multiple isoforms due to alternative splicing. The JNK1 protein is activated by dual phosphorylation on threonine and tyrosine residues a process integral to its function as a stress-activated protein kinase. Commonly used research tools include JNK antibodies and phospho JNK antibodies which help detect the activated forms of this kinase during cellular studies.
- Biological function summary
This kinase plays an important role in processes such as inflammation apoptosis and cellular stress responses. JNK1 is not just an isolated enzyme. It forms complexes with other proteins under specific conditions facilitating diverse cellular responses. For example JNK1 activation influences transcription factors like c-Jun by phosphorylating them impacting gene expression related to cell survival and death. This activity establishes JNK1 as a significant player in routine cell functioning and response to external environmental stressors.
- Pathways
The kinase is part of the MAPK signaling pathways and the stress-activated protein kinase (SAPK) pathways. These pathways involve multiple signaling cascades important for transmitting extracellular signals into the cellular environment. JNK1 interacts with proteins like MKK4 and MKK7 which are upstream activators and ATF2 a downstream target. This positioning makes JNK1 an essential signaling node that translates extracellular stressors into cellular responses providing adaptability to cells amidst changing conditions.
- Associated diseases and disorders
JNK1 has connections to disorders such as cancer and neurodegenerative diseases. Abnormal activation of JNK1 can lead to irregular cell proliferation making it pertinent in cancer. Similarly in neurodegenerative diseases like Alzheimer's JNK1's involvement in neuronal apoptosis turns critical. It regulates tau phosphorylation connecting JNK1 with tauopathies observed in Alzheimer's disease. Researchers often investigate these associations to understand the mechanisms that underpin these conditions and develop targeted therapies.
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3 product images
Next Generation Sequencing - Human MAPK8 knockout U-2 OS cell line (ab277181)
1 bp insertion after Pro59 of the WT protein
Western blot - Human MAPK8 knockout U-2 OS cell line (ab277181)
False colour image of Western blot: Anti-JNK1 antibody [EPR17557] staining at 1/2500 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-JNK1 antibody [EPR17557] ab199380 was shown to bind specifically to JNK1. A band was observed at 42/48 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in mapk8 knockout cell line ab277181 (knockout cell lysate ab277223). To generate this image, wild-type and mapk8 knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-JNK1 antibody [EPR17557] (Anti-JNK1 antibody [EPR17557] ab199380) at 1/2500 dilution
Lane 1: Wild-type U-2 OS cell lysate at 20 µg
Lane 2: MAPK8 knockout U-2 OS cell lysate at 20 µg
Performed under reducing conditions.
Observed band size: 42-48 kDa
Western blot - Human MAPK8 knockout U-2 OS cell line (ab277181)
False colour image of Western blot: Anti-JNK1 antibody [EPR140(2)] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-JNK1 antibody [EPR140(2)] ab110724 was shown to bind specifically to JNK1. A band was observed at 42/48 kDa in wild-type U-2 OS cell lysates with no signal observed at this size in mapk8 knockout cell line ab277181 (knockout cell lysate ab277223). To generate this image, wild-type and mapk8 knockout U-2 OS cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-JNK1 antibody [EPR140(2)] (Anti-JNK1 antibody [EPR140(2)] ab110724) at 1/1000 dilution
Lane 1: Wild-type U-2 OS cell lysate at 20 µg
Lane 2: MAPK8 knockout U-2 OS cell lysate at 20 µg
Performed under reducing conditions.
Observed band size: 42-48 kDa
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