Phospho-CAD (Ser1859) (D5K5W) Rabbit mAb #67235

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Supporting Data

REACTIVITY	H M R
SENSITIVITY	Endogenous
MW (kDa)	240
Source/Isotype	Rabbit IgG

Application Key:

WB-Western Blotting
IHC-Immunohistochemistry

Species Cross-Reactivity Key:

H-Human
M-Mouse
R-Rat

Product Information

Product Usage Information

Application	Dilution
Western Blotting	1:1000
Immunohistochemistry (Paraffin)	1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Protocol

Specificity / Sensitivity

Phospho-CAD (Ser1859) (D5K5W) Rabbit mAb recognizes endogenous levels of CAD protein only when phosphorylated at Ser1859.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser1859 of human CAD protein.

Background

CAD is essential for the de novo synthesis of pyrimidine nucleotides and possesses the following enzymatic activities: glutamine amidotransferase, carbamoyl-phosphate synthetase, aspartate transcarbamoylase, and dihydroorotase. Thus, the enzyme converts glutamine to uridine monophosphate, a common precursor of all pyrimidine bases, and it is necessary for nucleic acid synthesis (1). In resting cells, CAD is localized mainly in the cytoplasm where it carries out pyrimidine synthesis. As proliferating cells enter S phase, MAP Kinase (Erk1/2) phosphorlyates CAD at Thr456, resulting in CAD translocation to the nucleus. As cells exit S phase, CAD is dephosphorylated at Thr456 and phosphorylated at Ser1406 by PKA, returning the pathway to basal activity (2). Various research studies have shown increased expression of CAD in several types of cancer, prompting the development of pharmacological inhibitors such as PALA. Further studies have identified CAD as a potential predictive early marker of prostate cancer relapse (3).
mTORC1 is a protein kinase that works to regulate the growth and proliferation of cells by sensing and integrating various growth signals. S6 kinase 1 (S6K1) is a downstream ribosomal protein target of mTORC1 and directly phosphorylates Ser1859 on CAD. This phosphorylation stimulates the first three steps of the de novo pyrimidine synthesis and thus helps to advance the cells overall progression through S phase of the cell cycle (4,5).

Database Links

UniProt ID: P27708

Entrez-Gene Id: 790

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