Product Name:
CAD-pS1859
Product Number:
ab-pn691
Target Full Name: Aspartate carbamoyltransferase
Target Alias: Carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase; EC 2.1.3.2; EC 3.5.2.3; EC 6.3.5.5; PYR1
Product Type Specific: CAD phosphosite-specific antibody
Antibody Code: PN691
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S1859
Protein UniProt: P27708
Protein SigNET: CAD
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human CAD
Antibody Immunogen Sequence: HRA(pS)DPG(βA)C
Antibody Immunogen Description: Corresponds to amino acid residues H1856 to G1862. S1859 phosphorylation stimulates enzyme activity. This is the major in vivo phosphorylation site in CAD (≥162 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). However, This human phosphosite is not highly conserved in nature, but is found in the rhesus monkey. CAD is known to be phosphorylated at this site in vitro by PKACa (PRKACA), and p70 S6Kb (RPS6KB1).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.5 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rat | Mouse | Hamster.
Scientific Background: CAD (Multifunctional protein CAD) is a multi-enzyme complex involved in pyrimidine nucleotide biosynthesis. This protein is a fusion protein encoding four enzymatic activities of the pyrimidine pathway. The first 3 enzymatic activities of the de novo pyrimidine pathway: carbamoylphosphate synthetase (CPSase; EC 6.3.5.5), aspartate transcarbamylase (ATCase; EC 2.1.3.2) and dihydroorotase (DHOase; EC 3.5.2.3). The CPSase-function is accomplished in 2 steps, by a glutamine-dependent amidotransferase activity (GATase) that binds and cleaves glutamine to produce ammonia, followed by an ammonium-dependent carbamoyl phosphate synthetase, which reacts with the ammonia, hydrogencarbonate and ATP to form carbamoyl phosphate. The endogenously produced carbamoyl phosphate is sequestered and channeled to the ATCase active site. ATCase then catalyzes the formation of carbamoyl-L-aspartate from L-aspartate and carbamoyl phosphate. In the last step, DHOase catalyzes the cyclization of carbamoyl aspartate to dihydroorotate. GATase (glutamine amidotransferase) and CPSase (carbamoyl phosphate synthase) form together the glutamine-dependent CPSase (GD-CPSase) (EC 6.3.5.5). In the central section; belongs to the metallo-dependent hydrolases superfamily DHOase family, CAD subfamily. CAD is directly phosphorylated by S6 kinase (a downstream target of mTORC1), regulating cell proliferation. CAD activity is allosterically inhibited by UTP (end product) and activated by PRPP (5-phosphoribosyl-1-pyrophosphate). CAD moves between the cytosol and nucleus; its presence in the nucleus promotes high-rate pyrimidine biosynthesis. CAD self-assembles into large hexamers, which are essential for the metabolic channelling of intermediates between domains. CAD is crucial for cellular growth and is often upregulated in cancer cells to meet high pyrimidine demands. This description may include information annotated by UniProt and/or Google AI.