CPS1 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 insertion in exon 15 and Insertion of the selection cassette in exon 15.
CPSM_HUMAN, CPSase 1, CPSase I, Carbamoyl phosphate synthase, Carbamoyl phosphate synthase [ammonia] mitochondrial, Carbamoyl phosphate synthetase 1, Carbamoyl phosphate synthetase 1 mitochondrial, Carbamoyl-phosphate synthase [ammonia], Carbamoyl-phosphate synthetase I, MS738, mitochondrial
CPS1 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 insertion in exon 15 and Insertion of the selection cassette in exon 15.
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.
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.
CPS1 also known as carbamoyl-phosphate synthetase 1 is an important mitochondrial enzyme with a mass of around 165 kDa. It acts in the urea cycle by converting ammonia and bicarbonate into carbamoyl phosphate. CPS1 expression occurs in the liver and intestinal enterocytes which situates it in tissues responsible for metabolism and detoxification processes. This enzyme plays a critical role in the removal of excess nitrogen from the body.
Carbamoyl-phosphate synthetase 1 functions as an initial step enzyme in the urea cycle which is a metabolic pathway for nitrogen disposal. It does not function alone as it forms part of multi-enzyme complexes to enhance efficiency in the cycle. These complexes facilitate the process of converting toxic ammonia into urea which the body can safely excrete. The proper function of CPS1 ensures that ammonia levels remain non-toxic.
The urea cycle remains the chief pathway involving CPS1 collaborating with enzymes like ornithine transcarbamylase and argininosuccinate synthetase. This cycle intersects with the mitochondrial respiratory chain showcasing a relationship between CPS1 and cellular energy balance. Besides the urea cycle the enzyme also links to other metabolic functions like gluconeogenesis given its prerequisite for maintaining a stable nitrogen homeostasis.
CPS1 deficiencies can lead to urea cycle disorders like hyperammonemia characterized by elevated ammonia levels in blood. This may result in neurological damage if untreated. Another condition related to CPS-1 is pulmonary arterial hypertension where studies suggest mishandling of ammonia may alter blood pressure regulation. Genetic mutations affecting the CPS1 protein often underlie these conditions highlighting the importance of genetic screening in affected individuals.
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Lanes 1-3: Merged signal (red and green). Green - Anti-CPS1 antibody [EPR7493-29] ab155083 observed at 165 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-CPS1 antibody [EPR7493-29] ab155083 Anti-CPS1 antibody [EPR7493-29] was shown to specifically react with CPS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261809 (knockout cell lysate Human CPS1 knockout HeLa cell lysate ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. Anti-CPS1 antibody [EPR7493-29] ab155083 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CPS1 antibody [EPR7493-29] (Anti-CPS1 antibody [EPR7493-29] ab155083) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CPS1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human CPS1 knockout HeLa cell line (ab261809)
Lane 3: Human liver tissue lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 165 kDa
Observed band size: 165 kDa
Lanes 1-3: Merged signal (red and green). Green - Anti-CPS1 antibody [EPR7493-3] ab129076 observed at 165 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-CPS1 antibody [EPR7493-3] ab129076 Anti-CPS1 antibody [EPR7493-3] was shown to specifically react with CPS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261809 (knockout cell lysate Human CPS1 knockout HeLa cell lysate ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. Anti-CPS1 antibody [EPR7493-3] ab129076 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CPS1 antibody [EPR7493-3] (Anti-CPS1 antibody [EPR7493-3] ab129076) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CPS1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human CPS1 knockout HeLa cell line (ab261809)
Lane 3: Human liver tissue lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 165 kDa
Observed band size: 165 kDa
Allele-2: Insertion of the selection cassette in exon 15.
Allele-1: 1 bp insertion in exon 15.
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