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AB264722

Human MC2R knockout HeLa cell line

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MC2R KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2.

View Alternative Names

ACTH receptor, ACTHR_HUMAN, Adrenocorticotropic hormone receptor, Adrenocorticotropin receptor, Corticotropin receptor, MC2 receptor, MC2-R, Melanocortin 2 receptor, Melanocortin 2 receptor (adrenocorticotropic hormone), Melanocortin receptor 2

3 Images
Sanger Sequencing - Human MC2R knockout HeLa cell line (AB264722)
  • Sanger seq

Unknown

Sanger Sequencing - Human MC2R knockout HeLa cell line (AB264722)

Allele-1 : 1 bp deletion in exon 2.

Sanger Sequencing - Human MC2R knockout HeLa cell line (AB264722)
  • Sanger seq

Lab

Sanger Sequencing - Human MC2R knockout HeLa cell line (AB264722)

Sequencing chromatogram displaying sequence edit in exon 2

Sanger Sequencing - Human MC2R knockout HeLa cell line (AB264722)
  • Sanger seq

Unknown

Sanger Sequencing - Human MC2R knockout HeLa cell line (AB264722)

Allele-2 : 1 bp insertion in exon 2.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 1 bp insertion in exon 2

Disease

Adenocarcinoma

Product details

Recommended control: Human wild-type HeLa cell line (ab255928). 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, 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?

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Properties and storage information

Gene name
MC2R
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

The melanocortin 2 receptor (MC2-R) also known as ACTH receptor is a protein that mechanically functions as a G protein-coupled receptor (GPCR). It is responsible for mediating activity through adenylate cyclase signaling by binding with adrenocorticotropic hormone (ACTH). This receptor has a molecular mass of approximately 33 kDa. MC2-R is mostly expressed in the adrenal cortex where it plays a role in synthesizing and releasing glucocorticoids.
Biological function summary

The action of MC2-R initiates the adrenal response to stress through glucocorticoid production which is important for maintaining homeostasis in the body. MC2-R functions as a standalone entity rather than as part of a larger complex. By binding to ACTH MC2-R triggers downstream signaling that leads to steroidogenesis. The biosynthesis of glucocorticoids helps in managing energy immune function and circadian rhythm.

Pathways

MC2-R is integral in the cAMP signaling pathway and the stress response pathway. In the cAMP signaling activation of MC2-R by ACTH results in increased production of cyclic AMP further promoting enzyme activation essential for glucocorticoid synthesis. Its activity is connected to the broader steroid hormone biosynthesis pathway working closely with proteins like StAR (steroidogenic acute regulatory protein) which facilitates cholesterol transport to the inner mitochondrial membrane.

Disruptions in MC2-R function can lead to conditions like familial glucocorticoid deficiency and Cushing's syndrome. Dysfunctional receptor signaling may lead to inadequate glucocorticoid production or excessive hormone levels. The role of MC2-R in these disorders connects it closely with proteins like MC1-R another melanocortin receptor involved in pigmentation and inflammatory response which also signifies related signaling cascade abnormalities.

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 protocols

Product promise

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