Human CPS1 knockout HeLa cell line
- Advanced Validation
- What is this?
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CPS1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and Insertion of the selection cassette in exon 15. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
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
- WB
Lab
Western blot - Human CPS1 knockout HeLa cell line (AB261809)
Lanes 1-3 : Merged signal (red and green). Green - ab155083 observed at 165 kDa. Red - loading control, ab8245 observed at 37 kDa.
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 ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. ab155083 and 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 (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/cps1-antibody-epr7493-29-ab155083'>ab155083</a>) 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
Predicted band size: 165 kDa
Observed band size: 165 kDa
false
- WB
Lab
Western blot - Human CPS1 knockout HeLa cell line (AB261809)
Lanes 1-3 : Merged signal (red and green). Green - ab129076 observed at 165 kDa. Red - loading control, ab8245 observed at 37 kDa.
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 ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. ab129076 and 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 (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/cps1-antibody-epr7493-3-ab129076'>ab129076</a>) 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
Predicted band size: 165 kDa
Observed band size: 165 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CPS1 knockout HeLa cell line (AB261809)
Allele-2 : Insertion of the selection cassette in exon 15.
- Sanger seq
Unknown
Sanger Sequencing - Human CPS1 knockout HeLa cell line (AB261809)
Allele-1 : 1 bp insertion in exon 15.
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
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.
Pathways
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.
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
Complementary Products
Primary Antibodies
AB129076
Anti-CPS1 antibody [EPR7493-3]
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPS1 antibody [EPR7493-3] (AB129076)](https://content.abcam.com/products/images/cps1-antibody-epr7493-3-ab129076--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img129382.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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