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Purified anti-CPS1 Antibody

Pricing & Availability
Clone
W21062G (See other available formats)
Regulatory Status
RUO
Other Names
Carbamoyl-Phosphate Synthetase I, Carbamoyl-Phosphate Synthase 1, Carbamoyl-Phosphate Synthase 1, Mitochondrial, Carbamoyl-Phosphate Synthase [Ammonia], Mitochondrial, CPSase I, CPSase 1
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
a. W21062G_PURE_CPS1_WB_102023
Whole cell extracts (15 µg total protein) from indicated cell lines and tissue were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL of purified anti-CPS1 (clone W21062G) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • a. W21062G_PURE_CPS1_WB_102023
    Whole cell extracts (15 µg total protein) from indicated cell lines and tissue were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL of purified anti-CPS1 (clone W21062G) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-rat IgG (Cat. No. 405405). Direct-Blot™ HRP anti-β-actin (Cat. No. 664804) was used as a loading control at a 1:50000 dilution. Western-Ready™ ECL Substrate Premium Kit (Cat. No. 426319) was used as a detection agent. Lane M: Molecular weight marker
  • b. W21062G_PURE_CPS1_ICC-1_110723
    HeLa cells were stained with 500 nM of MitoSpy™ Red CMXRos (Cat. No. 424802) (panel A, pseudo color green) for 30 minutes at 37°C, fixed with Fixation Buffer (Cat. No. 420801) for 10 minutes and permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 10.0 µg/mL of purified anti-CPS1 (clone W21062G), followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) (panel B, magenta) for 1 hour at room temperature. Cells were also stained with Flash Phalloidin™ Green 488 (Cat. No. 424201) (panel C, pseudo color yellow) and nuclei were counterstained with DAPI (Cat. No. 422801) (panel D, blue). Images were merged (panel D). The images were captured by a Revvity Operetta CLS™ High Content Analysis System with a 63X objective. Scale bar: 50 µm
  • c. W21062G_PURE_CPS1_ICC-2_110723
    HeLa cells were stained with 500 nM of MitoSpy™ Red CMXRos (Cat. No. 424802) (panel A, pseudo color green) for 30 minutes at 37°C, fixed and permeabilized with methanol for 10 minutes, and blocked with 5% FBS for 1 hour at room temperature. Cells were then stained with 5.0 µg/mL of purified anti-CPS1 (clone W21062G), followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) (panel B, magenta) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801) (panel C, blue). Images were merged (panel D). The images were captured by a Revvity Operetta CLS™ High Content Analysis System with a 63X objective. Scale bar: 50 µm
  • d. W21062G_PURE_CPS1_IHC-P_110723
    IHC staining of purified anti-CPS1 (clone W21062G) on formalin-fixed paraffin-embedded human liver tissue. Following antigen retrieval using Tris-EDTA (10 mM Tris, 1 mM EDTA, pH 9.0), the tissue was incubated with 0.5 µg/mL of purified anti-CPS1 (clone W21062G) followed by incubation with Alexa Fluor® 647 Goat anti-rat IgG (Cat. No. 405416) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 10X (panel A) or 40X objective (panel B). Scale bar: 50 µm
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630751 25 µg £113
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630752 100 µg £284
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Description

CPS1, or carbamoyl phosphate synthase 1, catalyzes the initial step in the urea cycle to produce carbamoyl phosphate from ammonia and bicarbonate. CPS1 is highly expressed in the mitochondria of hepatocytes, where a robust urea cycle takes place to detoxify ammonia from protein catabolism. CPS1 deficiency in humans is an autosomal recessive disease that causes severe symptoms, such as hyperammonemia, in urea cycle disorders. This deficiency affects the neuronal and metabolic development of newborn children. CPS1 undergoes various post-translational modifications, including acetylation, succinylation, and glutarylation, which regulate the enzymatic activity of CPS1. During fasting, sirtuin 5 (SIRT5) is activated by increased NAD levels, leading to the deacetylation and deglutarylation of CPS1. This process removes inhibitory modifications from the active ATP binding site. The tumor suppressor p53 regulates ammonia metabolism by repressing the urea cycle and the expression of genes such as CPS1, OTC, and ARG1. The repression leads to the inhibition of ureagenesis and the elimination of ammonia, ultimately impacting tumor growth.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Recombinant fragment of human CPS1
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IHC-P - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.25 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 10.0 μg/mL is recommended. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 0.25 - 1.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

For immunocytochemistry (ICC), the recommended fixation/permeabilization method is either 4% paraformaldehyde/0.5% Triton X-100, paraformaldehyde followed by ice-cold methanol, or ice-cold methanol fixed only.

For immunohistochemistry (IHC), the recommended antigen retrieval system is either Citrate Buffer, 10X (Cat. No. 420902) or Tris-EDTA pH 9.0 buffer.

RRID
AB_3083419 (BioLegend Cat. No. 630751)
AB_3083419 (BioLegend Cat. No. 630752)

Antigen Details

Structure
3 isoforms exist/form homodimer and homooligomer
Distribution

Liver and small intestine/mitochondria

Function
Catalyzes the urea cycle of ureotelic animals and remove excess ammonia from the cell
Interaction
Post-translational modification by SIRT5/Catalyze ammonia and bicarbonate to carbamoyl phosphate
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Proliferation and Viability, Mitochondrial Function, Neuroscience
Molecular Family
Enzymes and Regulators, Mitochondrial Markers
Antigen References
  1. Nakagawa T, et al. 2009. Cell. 137(3):560-70.
  2. Diez-Fernandez C, et al. 2017. Expert Opin. Ther. Targets. 21:391-399.
  3. Li L, et al 2019. Nature. 567:253-256. 
  4. Nitzahn M, et al. 2020. Mol Genet Metab. 131:289-298.
Gene ID
1373 View all products for this Gene ID
UniProt
View information about CPS1 on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 1    Revision Date: 10/20/2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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