MEF2C 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: 65 bp deletion in Exon 4 and intron 2-3.
Images
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 65 bp deletion in Exon 4 and intron 2-3.
Recommended products
MEF2C 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: 65 bp deletion in Exon 4 and intron 2-3.
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 65 bp deletion in Exon 4 and intron 2-3.
- Disease
- Acute Monocytic Leukemia
- Concentration
Properties
- Gene name
- MEF2C
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- VWA, TPOX, TH01, D7S820, D5S818, D16S539, CSF1PO, D13S317
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Suspension
- 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
- Culture medium
- RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
- 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
**Recommended control:** Human wild-type THP-1 cell line (ab281894). 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. **Cryopreservation cell medium: **Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose. **Culture medium: **RPMI + 10% FBS + 0.05 mM β-mercaptoethanol **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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2-4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. 4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily. 5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL. **Subculture guidelines:**
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
The MEF2C protein also known as MADS box transcription enhancer factor 2 polypeptide C plays an important role in gene expression regulation. This protein has a mass of approximately 52 kDa and is mainly found in the heart skeletal muscle brain and endothelial cells. It is part of the MEF2 family which includes other members like MEF2A MEF2B and MEF2D. MEF2C acts as a transcription factor binding to specific DNA sequences to activate the transcription of target genes involved in various biological processes.
- Biological function summary
The MEF2C protein is essential in cellular differentiation and development. MEF2C is not part of a complex but interacts with other cofactors and regulatory proteins to fulfill its roles. It is important in the development of cardiac and skeletal muscle cells and plays a significant role in neurogenesis and neuronal connectivity in the central nervous system. This protein's precise regulation ensures proper development and function of these tissues and organs.
- Pathways
MEF2C influences cellular signaling through the MAPK/ERK and calcium/calmodulin-dependent protein kinase pathways. In the MAPK/ERK pathway MEF2C modulates gene expression in response to external stimuli interacting with proteins like ERK1/2. In the calcium/calmodulin-dependent kinase pathway MEF2C plays a role in calcium signaling networks impacting neuronal plasticity and survival. These pathways highlight MEF2C’s involvement in muscle and brain function.
- Associated diseases and disorders
MEF2C mutations or dysregulation link to conditions like dilated cardiomyopathy and neurodevelopmental disorders such as MEF2C haploinsufficiency syndrome. In cardiac disease abnormal MEF2C activity can affect heart muscle cell function often involving proteins like actin and myosin. In neurodevelopmental disorders MEF2C's interaction with various transcriptional regulators impacts neuronal networks contributing to symptoms like impaired cognitive development and autism spectrum-like behaviors.
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3 product images
Western blot - Human MEF2C knockout THP-1 cell line (ab313739)
False colour image of Western blot: Anti-MEF2C antibody [EPR19089-202] - ChIP Grade 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-MEF2C antibody [EPR19089-202] - ChIP Grade ab211493 was shown to bind specifically to MEF2C. A band was observed at 55/60 kDa in wild-type THP-1 cell lysates with no signal observed at this size in MEF2C knockout cell line (ab313739). To generate this image, wild-type and MEF2C knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-MEF2C antibody [EPR19089-202] - ChIP Grade (Anti-MEF2C antibody [EPR19089-202] - ChIP Grade ab211493) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: MEF2C knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human MEF2C knockout THP-1 cell line (ab313739)
Lane 3: Daudi cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 55/60 kDa, 37 kDa
Western blot - Human MEF2C knockout THP-1 cell line (ab313739)
False colour image of Western blot: Anti-MEF2A + MEF2C antibody [EPR19089-34] 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-MEF2A + MEF2C antibody [EPR19089-34] ab197070 was shown to bind specifically to MEF2C. A band was observed at 55/60 kDa in wild-type THP-1 cell lysates with no signal observed at this size in MEF2C knockout cell line (ab313739). To generate this image, wild-type and MEF2C knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-MEF2A + MEF2C antibody [EPR19089-34] (Anti-MEF2A + MEF2C antibody [EPR19089-34] ab197070) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: MEF2C knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human MEF2C knockout THP-1 cell line (ab313739)
Lane 2: Western blot - Human MEF2C knockout THP-1 cell line (Human MEF2C knockout THP-1 cell line ab288702)
Lane 3: Daudi cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 55/60 kDa, 37 kDa
Sanger Sequencing - Human MEF2C knockout THP-1 cell line (ab313739)
65 bp deletion in Exon 4
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