Recombinant Anti-CAD antibody [EP710Y] (ab40800)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP710Y] to CAD
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CAD antibody [EP710Y]
See all CAD primary antibodies -
Description
Rabbit monoclonal [EP710Y] to CAD -
Host species
Rabbit -
Specificity
ab40800 is specific to the N-terminus of CAD.
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Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa and HEK293 cell lysate. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate (ab150035).
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP710Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab40800 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/120.
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ICC/IF |
1/100 - 1/300.
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WB |
1/1000. Predicted molecular weight: 243 kDa.
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IP |
1/30 - 1/50.
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Notes |
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Flow Cyt (Intra)
1/120. |
ICC/IF
1/100 - 1/300. |
WB
1/1000. Predicted molecular weight: 243 kDa. |
IP
1/30 - 1/50. |
Target
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Relevance
Carbamoyl phosphate synthetase-aspartate carbamoyltransferase-dihydroorotase (CAD) is a multifunctional protein that initiates and regulates mammalian de novo pyrimidine biosynthesis. This trifunctional protein which is associated with the enzymatic activities of the first 3 enzymes in the 6-step pathway of pyrimidine biosynthesis is the rate-limiting step in the de novo pyrimidine synthetic pathway. Although most of the CAD protein in the cell is cytosolic, phosphorylation at threonine 456 localizes the protein to the nucleus. While MAPK and EGF phosphorylate CAD at threonine 456, MAPK and c-myc have been found to induce over-expression of CAD. -
Cellular localization
Cytoplasmic and Nuclear -
Database links
- Entrez Gene: 790 Human
- Omim: 114010 Human
- SwissProt: P27708 Human
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Alternative names
- Aspartate carbamoyltransferase antibody
- Aspartate transcarbamylase antibody
- CAD protein antibody
see all
Images
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Anti-CAD antibody [EP710Y] (ab40800) at 1/2000 dilution (purified) + HeLa cell lysate at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 243 kDa
Observed band size: 250 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunofluorescence staining of HeLa cells with purified ab40800 at a working dilution of 1/300, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab40800 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling CAD (red) with ab40800 at a 1/120 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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ab40800 (purified) at 1/30 immunoprecipitating CAD in 10 μg HeLa (Lanes 1 and 2, observed at 250 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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Anti-CAD antibody [EP710Y] (ab40800) at 1/1000 dilution (purified) + HeLa cell lysate at 1/20000 dilution
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 243 kDa
Observed band size: 250 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Unpurified ab40800 (1/100), staining human CAD (N-term) in HeLa cells by Immunofluorescence.
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Anti-CAD antibody [EP710Y] (ab40800) at 1/1000 dilution (unpurified) +
HEK-293 whole cell lysate (ab7902) at 10 µg
Predicted band size: 243 kDa
Observed band size: 250 kDa why is the actual band size different from the predicted?
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab40800 has been referenced in 9 publications.
- Mehazri L et al. Loss of Fer Jeopardizes Metabolic Plasticity and Mitochondrial Homeostasis in Lung and Breast Carcinoma Cells. Int J Mol Sci 22:N/A (2021). PubMed: 33806191
- Verrier ER et al. Combined small molecule and loss-of-function screen uncovers estrogen receptor alpha and CAD as host factors for HDV infection and antiviral targets. Gut 69:158-167 (2020). WB ; Human . PubMed: 30833451
- Brandt J et al. The Cellular Protein CAD is Recruited into Ebola Virus Inclusion Bodies by the Nucleoprotein NP to Facilitate Genome Replication and Transcription. Cells 9:N/A (2020). PubMed: 32370067
- Pesini A et al. Brain pyrimidine nucleotide synthesis and Alzheimer disease. Aging (Albany NY) 11:8433-8462 (2019). PubMed: 31560653
- Lee JS et al. Urea Cycle Dysregulation Generates Clinically Relevant Genomic and Biochemical Signatures. Cell 174:1559-1570.e22 (2018). PubMed: 30100185
- Sun W et al. Interaction between von Hippel-Lindau Protein and Fatty Acid Synthase Modulates Hypoxia Target Gene Expression. Sci Rep 7:7190 (2017). PubMed: 28775317
- Rabinovich S et al. Diversion of aspartate in ASS1-deficient tumours fosters de novo pyrimidine synthesis. Nature 527:379-383 (2015). WB . PubMed: 26560030
- DeVito SR et al. Cytomegalovirus-mediated activation of pyrimidine biosynthesis drives UDP-sugar synthesis to support viral protein glycosylation. Proc Natl Acad Sci U S A 111:18019-24 (2014). PubMed: 25472841
- Alfonso-Pérez T et al. Cytoplasmic interaction of the tumour suppressor protein hSNF5 with dynamin-2 controls endocytosis. Oncogene N/A:N/A (2013). PubMed: 23851497