AB313739

Human MEF2C knockout THP-1 cell line

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MEF2C KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 65 bp deletion in Exon 4 and intron 2-3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

3 Images

Supplier Data

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] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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 (<a href='/en-us/products/primary-antibodies/mef2c-antibody-epr19089-202-chip-grade-ab211493'>ab211493</a>) 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

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 51 kDa

Observed band size: 55/60 kDa,37 kDa

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Supplier Data

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] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/mef2a-mef2c-antibody-epr19089-34-ab197070'>ab197070</a>) 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 (<a href='/en-us/products/cell-lines/human-mef2c-knockout-thp-1-cell-line-ab288702'>ab288702</a>)

Lane 3:

Daudi cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 55/60 kDa,37 kDa

false

Sanger Sequencing - Human MEF2C knockout THP-1 cell line (AB313739)

Sanger seq

Lab

Sanger Sequencing - Human MEF2C knockout THP-1 cell line (AB313739)

65 bp deletion in Exon 4

Key facts

Cell type

THP-1

Species or organism

Human

Tissue

Blood

Form

Liquid

form

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

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.

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What's included?

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Properties and storage information

Gene name

MEF2C

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

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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.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Quality control

STR analysis

VWA, TPOX, TH01, D7S820, D5S818, D16S539, CSF1PO, D13S317

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Target data

See full target information MEF2C

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

Human MEF2C knockout THP-1 cell line

Western blot - Human MEF2C knockout THP-1 cell line (AB313739)

Western blot - Human MEF2C knockout THP-1 cell line (AB313739)

Sanger Sequencing - Human MEF2C knockout THP-1 cell line (AB313739)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Disease

Reactivity data

Product details

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

Subculture guidelines

Culture medium

Cryopreservation medium

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Target data

Complementary Products

Cell Lines & Lysates

Primary Antibodies

Primary Antibodies

Primary Antibodies

Product promise