NPR3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 1 bp insertion in exon 1 and 2 bp deletion in exon 1.
NPR3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 1 bp insertion in exon 1 and 2 bp deletion in exon 1.
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.
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 HeLa cell line (ab255448). 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: DMEM (High Glucose) + 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. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10E4 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 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
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.
The NPR-C also known as natriuretic peptide receptor C functions as an important regulator in the homeostasis of cardiovascular and renal functions. It is also referred to as atrial natriuretic peptide receptor C (ANPRC). This protein does not possess guanylyl cyclase activity unlike its counterparts NPR-A and NPR-B but instead acts via clearance binding of natriuretic peptides preventing them from binding to other receptors. The receptor has a molecular mass of approximately 60 kDa and is widely expressed in tissues such as vascular endothelium kidney and adipose tissue.
NPR-C binds multiple natriuretic peptides including ANP BNP and CNP and modulates their effects by facilitating their clearance from circulation. This receptor does not function as part of a larger protein complex unlike other receptor subtypes. It controls blood volume and pressure by influencing the natriuretic diuretic and vasorelaxant actions of its peptide ligands. NPR-C helps maintain fluid balances and prevents overactivation of hormonal pathways linked to cardiovascular conditions.
NPR-C serves an important role in the natriuretic peptide signaling pathway primarily through the regulation of peptide availability. It physically interacts with several circulating natriuretic peptides but not through second messenger cascades. NPR-C is integral to maintaining the balance between the ANP receptor pathway and the renin-angiotensin-aldosterone system therefore indirectly modulating the functions of NPR-A and NPR-B which are responsible for cyclic GMP-dependent signaling.
NPR-C contributes to the pathophysiology of hypertension and heart failure by overseeing the peptide-hormone balance in the cardiovascular system. Anomalies in NPR-C expression or function can lead to improper regulation of blood pressure and fluid retention. Its interaction with proteins such as angiotensin II receptors can exacerbate the effects of hypertension. Understanding the role of NPR-C in these conditions presents potential therapeutic targets for modulating cardiovascular diseases.
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Full details and terms and conditions can be found here:
Terms & Conditions.
Allele-3: 1 bp insertion in exon 1.
Allele-1: 2 bp deletion in exon 1.
Allele-2: 1 bp deletion in exon 1.
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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