Human MAPK8 knockout U-2 OS cell line
Be the first to review this product! Submit a review
|
(0 Publication)
MAPK8 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion after Pro59 of the WT protein Frameshift: 100%. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View 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
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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)] (<a href='/en-us/products/primary-antibodies/jnk1-antibody-epr1402-ab110724'>ab110724</a>) 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
Observed band size: 42-48 kDa
false
- WB
Lab
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] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/jnk1-antibody-epr17557-ab199380'>ab199380</a>) 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
Observed band size: 42-48 kDa
false
- NGS
Lab
Next Generation Sequencing - Human MAPK8 knockout U-2 OS cell line (AB277181)
1 bp insertion after Pro59 of the WT protein
Product details
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 duration
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB179461
Anti-JNK1 + JNK2 + JNK3 antibody [EPR16797-211]
20ul selling size|RabMAb|Recombinant
![Flow Cytometry (Intracellular) - Anti-JNK1 + JNK2 + JNK3 antibody [EPR16797-211] (AB179461)](https://content.abcam.com/products/images/jnk1-jnk2-jnk3-antibody-epr16797-211-ab179461--flow-cytometry-intracellular-img25613.jpg)
- 4
- 8
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