MED12 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
Alternative names
ARC240, Activator-recruited cofactor 240 kDa component, CAG repeat protein 45, CAGH45, HOPA, KIAA0192, MED12_HUMAN, Mediator complex subunit 12, Mediator of RNA polymerase II transcription subunit 12, OPA-containing protein, TNRC11, Thyroid hormone receptor-associated protein complex 230 kDa component, Trap230, Trinucleotide repeat-containing gene 11 protein
Recommended products
MED12 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- HCT116
- Species or organism
- Human
- Tissue
- Colon
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- MED12
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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 conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HCT116 cell line (ab288559). 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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
MED12 also known as mediator complex subunit 12 is a component of the transcriptional Mediator complex. It weighs approximately 250 kDa and is widely expressed in many tissues showing high transcriptional regulatory activity. MED12 collaborates with other subunits within the Mediator complex to transmit signals from regulatory DNA elements to RNA polymerase II facilitating gene expression regulation.
- Biological function summary
MED12 functions as a core component of the larger Mediator complex which plays an important role in controlling RNA polymerase II-dependent genes. Through interactions with activators and repressors MED12 participates in chromatin modification and looping affecting gene transcription rates. It is enriched in tissues where fine control of gene expression is essential impacting growth differentiation and development processes.
- Pathways
MED12 integrates into critical signaling pathways including the Wnt/β-catenin and Hedgehog pathways. In these pathways MED12 interacts with other proteins like β-catenin serving as a connecting node that influences the transcription of target genes essential for cell proliferation and differentiation. Such interactions help coordinate cellular responses to external signals and ensure proper pathway regulation.
- Associated diseases and disorders
MED12 mutations have been linked to disorders like Lujan-Fryns syndrome and uterine leiomyomas. In these contexts MED12 interacts with G protein-coupled receptors and other transcription factors such as β-catenin resulting in dysregulation that leads to the development of these conditions. Understanding the connections between MED12 and associated proteins provides insights into the underlying mechanisms of these pathologies.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
2 product images
Western blot - Human MED12 knockout HCT116 cell line (ab286465)
Western blot: Anti-MED12 antibody [BLR084G] (Anti-MED12 antibody [BLR084G] ab275962) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-MED12 antibody [BLR084G] ab275962 was shown to bind specifically to MED12. A band was observed at 238-243 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in MED12 knockout cell line. To generate this image, wild-type and MED12 knockout HCT 116 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-MED12 antibody [BLR084G] (Anti-MED12 antibody [BLR084G] ab275962) at 1/1000 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: MED12 knockout HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human MED12 knockout HCT116 cell line (ab286465)
Lane 3: HeLa cell lysate at 20 µg
Lane 4: A549 Membrane cell lysate at 20 µg
SecondaryLanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 238-243 kDa
Next Generation Sequencing - Human MED12 knockout HCT116 cell line (ab286465)
118 bp deletion after Lys192
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
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