CAD Recombinant Monoclonal Antibody

Code CSB-RA909912A0HU
Size US$210
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  • Overlay Peak curve showing Hela cells stained with CSB-RA909912A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*106cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/1*106cells) used under the same conditions. Acquisition of >10, 000 events was performed.
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Product Details

Uniprot No.
Target Names
CAD
Alternative Names
CAD antibody; CAD protein [Includes: Glutamine-dependent carbamoyl-phosphate synthase antibody; EC 6.3.5.5); Aspartate carbamoyltransferase antibody; EC 2.1.3.2); Dihydroorotase antibody; EC 3.5.2.3)] antibody
Species Reactivity
Human
Immunogen
A synthesized peptide from human CAD protein
Immunogen Species
Homo sapiens (Human)
Conjugate
Non-conjugated
Clonality
Monoclonal
Isotype
Rabbit IgG
Clone No.
9E8
Purification Method
Affinity-chromatography
Concentration
It differs from different batches. Please contact us to confirm it.
Buffer
Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.
Form
Liquid
Tested Applications
ELISA, FC
Recommended Dilution
Application Recommended Dilution
FC 1:50-1:200
Troubleshooting and FAQs
Storage
Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
Lead Time
Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
Description

CAD represents a fascinating example of metabolic efficiency in mammalian cells, encoding a large trifunctional enzyme that catalyzes the first three steps of de novo pyrimidine biosynthesis. This multidomain protein combines glutamine-dependent carbamoyl-phosphate synthase, aspartate carbamoyltransferase, and dihydroorotase activities within a single polypeptide chain. Given pyrimidine nucleotides' essential roles in DNA and RNA synthesis, CAD activity is particularly elevated in rapidly proliferating cells, making it a compelling target for researchers investigating cell cycle regulation, cancer metabolism, and nucleotide biosynthesis pathways.

This recombinant monoclonal antibody, clone 9E8, offers the reproducibility and consistency that demanding experimental workflows require. Developed using recombinant technology with a defined sequence, this rabbit IgG eliminates the lot-to-lot variability that can compromise longitudinal studies or multi-site collaborations. The antibody was raised against a synthesized peptide derived from human CAD protein, ensuring precise epitope targeting.

Flow cytometry validation demonstrates clear performance in detecting intracellular CAD in HeLa cells. Using fixed and permeabilized cells, the antibody produces a distinct positive shift compared to isotype control, confirming specific detection of endogenous CAD protein. The validated protocol employed formaldehyde fixation with Triton X-100 permeabilization, providing researchers with an established starting point for their own intracellular staining experiments.

For investigators studying metabolic reprogramming in cancer, nucleotide pool regulation, or the molecular mechanisms governing cell proliferation, this antibody provides a reliable tool for flow cytometric analysis of CAD expression at the single-cell level. The unconjugated format allows flexibility in secondary antibody selection to accommodate various instrument configurations and experimental designs.

Usage
For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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Target Background

Function
This protein is a 'fusion' protein encoding four enzymatic activities of the pyrimidine pathway (GATase, CPSase, ATCase and DHOase).
Gene References into Functions
  1. CAD dihydroorotase and aspartate transcarbamoylase domains self-assembles into dimers and trimers. PMID: 28552578
  2. Detection of the CAD-ALK gene fusion in urine tr-DNA anticipated radiological confirmation of disease progression. Analysis of plasma ctDNA identified ALK kinase mutations that emerged during treatment with the ALK inhibitor entrectinib PMID: 28368455
  3. Charge neutralization in the active site of the catalytic trimer of aspartate transcarbamoylase promotes diverse structural changes. PMID: 28833948
  4. This study showed that CAD deficiency co-occurrence of anaemia, anisopoikilocytosis, global developmental delay, and seizures. PMID: 28007989
  5. Previous study found three metal ions in huDHOase active site; in the present study, the putative third metal binding site in type II enzymes, such as StDHOase and ScDHOase, was created and identified. PMID: 26446564
  6. Changes in glycosylation in caused by mutations in CAD. PMID: 25678555
  7. These results establish CAD as a downstream effector of Rheb and suggest a possible role of Rheb in regulating de novo pyrimidine nucleotide synthesis PMID: 25422319
  8. Recombinant aspartate carbamoyltransferase domain from the CAD enzyme complex forms homotrimers in solution. PMID: 24316846
  9. The results obtained indicate that mLST8 bridges between CAD and mTOR, and plays a role in the signaling mechanism where CAD is regulated in the mTOR pathway through the association with mLST8 PMID: 23594158
  10. preliminary X-ray diffraction analysis of the dihydroorotase domain of human CAD PMID: 23143245
  11. findings show that in prostate tumor cells, CAD fosters androgen receptor translocation into the nucleus and stimulates its transcriptional activity; in radical prostatectomy specimens, CAD expression was not correlated with proliferation markers, but a higher CAD mRNA level was associated with local tumor extension and cancer relapse PMID: 21982950
  12. the cad gene is regulated by a nonclassical ERalpha/Sp1-mediated pathway. PMID: 12746293
  13. Data show that PRMT5 can be found in association with hSWI/SNF complexes and is involved in regulating the expression of carbamoyl-phosphate synthase-aspartate carbamoyltransferase-dihydroorotase. PMID: 14559996
  14. the nuclear import of CAD appears to promote optimal cell growth PMID: 15890648
  15. cad gene expression is repressed by hypoxia-inducible factor-1alpha, which represents a functional link between hypoxia and cell-cycle arrest. PMID: 16155188
  16. hCPS_DeltaA was not able to fully assume the catalytically competent conformation, with specific activity of CP formation decreased 700-fold. PMID: 18679823

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Involvement in disease
Epileptic encephalopathy, early infantile, 50 (EIEE50)
Subcellular Location
Cytoplasm. Nucleus. Note=Cytosolic and unphosphorylated in resting cells, translocates to the nucleus in response to EGF stimulation, nuclear import promotes optimal cell growth.
Protein Families
Metallo-dependent hydrolases superfamily, DHOase family, CAD subfamily
Database Links

HGNC: 1424

OMIM: 114010

KEGG: hsa:790

STRING: 9606.ENSP00000264705

UniGene: Hs.377010

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No. 269, Shendun 5th Road, Donghu Hi-Tech Development Area, Hubei Province, 430206, P.R.China
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