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NOS3 KO cell line available to order. Free of charge wild type control provided.

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Key facts

Cell type

MCF7

Species or organism

Human

Tissue

Breast

Form

Liquid

Alternative names

Recommended products

NOS3 KO cell line available to order. Free of charge wild type control provided.

Key facts

Cell type

MCF7

Form

Liquid

Disease

Adenocarcinoma

Concentration
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Properties

Gene name

NOS3

Gene editing type

Knockout

Gene editing method

CRISPR technology

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Female

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

  • Slow to trypsinise.

  • All seeding densities should be based on cell counts gained by established methods.

  • A guide seeding density of 5-7x104 cells/cm2 is recommended.

  • Cells should be passaged when they have achieved 80-90% confluence.

Culture medium

MEM + 10% FBS + 0.01 mg/ml bovine insulin

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 MCF7 cell line (Human wild-type MCF7 cell line ab277916). 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

ENOS also known as endothelial nitric oxide synthase is an enzyme important for the production of nitric oxide (NO) in blood vessels. This protein with a molecular weight of approximately 133 kDa is expressed mostly in endothelial cells. eNOS plays a mechanical role in synthesizing NO from L-arginine a process requiring cofactors such as NADPH and oxygen. The activity of eNOS can be investigated through techniques such as Western blot with specific assays like phospho-eNOS ELISA available to measure its phosphorylated forms indicating activated states of the enzyme.

Biological function summary

ENOS contributes significantly to the regulation of vascular tone and blood flow by promoting vasodilation. It does not function alone; instead it forms complexes with other proteins to exert its full effect. eNOS activity influences the process of angiogenesis inflammation modulation and platelet aggregation. Through its ability to produce nitric oxide eNOS acts as a signaling molecule helping maintain vascular homeostasis.

Pathways

ENOS is a critical player in the NO signaling pathway and interacts intricately with the PI3K/Akt pathway. Nitric oxide produced by eNOS has a role in signaling cascades that lead to vascular dilation and reduced blood pressure. The PI3K/Akt pathway regulates eNOS activity via phosphorylation enhancing NO production. Other proteins like caveolin-1 and calmodulin modulate eNOS impacting these pathways' outcomes.

Associated diseases and disorders

ENOS is associated with cardiovascular conditions like atherosclerosis and hypertension. Dysfunction of eNOS can lead to reduced NO production impairing vasodilation and contributing to these diseases. In cardiovascular disorders eNOS interacts with proteins such as the angiotensin II type 1 receptor which can negatively impact its function exacerbating disease states. Investigating eNOS and its related proteins provides insight into potential therapeutic targets for improving cardiovascular health.

Product promise

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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:
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    Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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