PDE5A KO cell line available to order. Free of charge wild type control provided.
PDE5A KO cell line available to order. Free of charge wild type control provided.
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 U-87 MG cell line (ab278079). 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: EMEM + 10% FBS
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.
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.
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.
PDE5A also known as PDE5 or phosphodiesterase type 5 plays a critical role in hydrolyzing cyclic guanosine monophosphate (cGMP) to its inactive form. This enzyme is part of the phosphodiesterase family and has a molecular mass of approximately 100 kDa. PDE5A is expressed mainly in the smooth muscle cells of blood vessels and in the corpus cavernosum of the penis. Other expression sites include the lung heart and skeletal muscle.
PDE5A regulates intracellular levels of cGMP an important secondary messenger in various signaling pathways. It is not part of a large protein complex but directly influences the contractility of smooth muscle tissues. By controlling cGMP levels PDE5A modulates vascular tone and influences processes such as vasodilation. This regulation is essential for maintaining a balance in the contractile state of muscles impacting blood flow and other vital functions.
PDE5A contributes significantly to the nitric oxide (NO)-cGMP signaling pathway which is important for vascular relaxation. In this pathway nitric oxide stimulates guanylate cyclase to produce cGMP which then activates specific kinases and ion channels. Related proteins such as guanylate cyclase and cGMP-dependent protein kinases work alongside PDE5A to mediate smooth muscle relaxation and vasodilation particularly in penile erectile tissue.
PDE5A shows strong connections to erectile dysfunction and pulmonary hypertension. In the case of erectile dysfunction impaired regulation of cGMP by PDE5A affects the relaxation of smooth muscle leading to inadequate blood flow. Pulmonary hypertension involves elevated blood pressure in the lung arteries which PDE5A influences through its role in controlling vascular tone. The enzyme guanylate cyclase associates closely with PDE5A in these contexts as both influence cGMP pathways involved in these conditions.
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.
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