Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell line
- Advanced Validation
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- WB
Lab
Western blot - Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell line (AB265930)
Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : NPR1 knockout HeLa cell lysate 20 μg
Lane 3 : U-251 MG cell lysate 20 μg
Lane 4 : Human Heart tissue lysate 20 μg
Lanes 1 - 4 : Merged signal (red and green). Green - anti-NPR1 antibody observed at 140 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
anti-NPR1 antibody was shown to react with NPR1 in wild-type HeLa cells in Western blot with loss of signal observed in NPR1 knockout cell line ab265930 (NPR1 knockout cell lysate ab258080). Wild-type HeLa and NPR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with anti-NPR1 antibody and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
anti-NPR1 antibody at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-npr1-natriuretic-peptide-receptor-a-gc-a-knockout-hela-cell-lysate-ab258080'>ab258080</a>) at 20 µg
Lane 3:
U-251 MG cell lysate at 20 µg
Lane 4:
Human Heart tissue lysate at 20 µg
Secondary
Lanes 1 - 3:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lanes 1 - 3:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
false
- Sanger seq
Unknown
Sanger Sequencing - Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell line (AB265930)
Allele-1 : 5 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell line (AB265930)
Allele-2 : 1 bp insertion in exon 1.
Complementary Products
Cell Lines & Lysates
AB258080
Human NPR1 (Natriuretic Peptide Receptor A/GC-A) knockout HeLa cell lysate
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Reactivity data
Product details
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?
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
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.
Biological function summary
Natriuretic peptide receptor A plays a significant role in cardiovascular homeostasis. NPR-A is not part of a larger complex but works closely with other natriuretic peptide systems to regulate blood pressure electrolyte balance and fluid homeostasis. Its activation results in diuresis natriuresis and vasodilation contributing to its function in maintaining cardiovascular health. Efficient functioning of NPR-A is essential for modulating cardiovascular responses.
Pathways
NPR-A is important in the cyclic GMP pathway and the ANP signaling pathway. It interacts with proteins like NPR-C and soluble guanylyl cyclase modulating key physiological responses. NPR-A impacts smooth muscle relaxation and renal function by influencing cGMP levels and related downstream pathways. Its role within these pathways is fundamental to maintaining vascular tone and sodium excretion.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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