Human MAPK12 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human MAPK12 knockout HEK-293T cell line (AB266280)
Lanes 1-3 : Merged signal (red and green). Green - ab205926 observed at 42 kDa. Red - loading control ab8245 observed at 36 kDa.
ab205926 Anti-MAPK 12 antibody [EPR6528(N)] was shown to specifically react with MAPK 12 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266280 (knockout cell lysate ab258041) was used. Wild-type and MAPK 12 knockout samples were subjected to SDS-PAGE. ab205926 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 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-p38 gamma/MAPK12 antibody [EPR6528(N)] (<a href='/en-us/products/primary-antibodies/p38-gamma-mapk12-antibody-epr6528n-ab205926'>ab205926</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
MAPK12 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human MAPK12 knockout HEK-293T cell line (ab266280)
Lane 3:
Human skeletal muscle tissue lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human MAPK12 knockout HEK-293T cell line (AB266280)
Allele-2 : 1 bp insertion in exon 4.
- Cell Culture
Lab
Cell Culture - Human MAPK12 knockout HEK-293T cell line (AB266280)
Representative images MAPK12 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.
- Sanger seq
Unknown
Sanger Sequencing - Human MAPK12 knockout HEK-293T cell line (AB266280)
Allele-1 : 5 bp deletion in exon 4
Complementary Products
Primary Antibodies
AB205926
Anti-p38 gamma/MAPK12 antibody [EPR6528(N)]
RabMAb|Recombinant|KO Validated|Trial Size
![Immunocytochemistry/ Immunofluorescence - Anti-p38 gamma/MAPK12 antibody [EPR6528(N)] (AB205926)](https://content.abcam.com/products/images/p38-gamma-mapk12-antibody-epr6528n-ab205926--immunocytochemistry-immunofluorescence-img69322.jpg)
- 0
- 0
Cell Lines & Lysates
AB266355
Human MAPK9 (JNK2) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
Cell Lines & Lysates
AB266343
Human RAD23A (hHR23A) knockout HEK-293T cell line
Advanced Validation
- 0
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Reactivity data
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 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
DMEM (High Glucose) + 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
P38 gamma/MAPK12 influences the regulation of gene expression apoptosis and cell cycle processes. It operates within a specific MAP kinase signaling module that conveys signals from the cellular surface to the nucleus. This protein sometimes works in conjunction with other members of the MAPK family forming complexes that further fine-tune cellular responses under various conditions. Its activities help cells adapt to changes in their environment by modulating transcription factors and other important substrates.
Pathways
The MAPK pathway and the stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) pathway both involve p38 gamma/MAPK12. Within these pathways p38 gamma is closely related to proteins such as JNK1 and JNK2 which also respond to stress signals. These pathways are essential for transmitting signals that regulate cellular growth differentiation and stress responses revealing intricacies of cellular adaptation to the environment.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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