CART Antibody

R&D Systems | Catalog # AF163

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry, Immunocytochemistry, Electrophysiology

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CART Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CART
Gln28-Leu116
Accession # Q16568

Specificity

Detects human CART in direct ELISAs and Western blots. Also recognizes a truncated recombinant human CART containing amino acids 69‑116.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for CART Antibody

CART antibody in Human Brain Hippocampus Tissue by Immunohistochemistry (IHC-P).

Detection of CART in Human Brain Hypothalamus Tissue.

CART was detected in immersion fixed paraffin-embedded sections of human brain (Hypothalamus) tissue using Goat Anti-Human/Mouse/Rat CART Antigen Affinity-purified Polyclonal Antibody (Catalog # AF163) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to neurons. Staining was performed using our IHC Staining with VisUCyte HRP Polymer Detection Reagents.
CART antibody in Mouse Brain Tissue by Immunohistochemistry (IHC-Fr).

CART in Mouse Brain Tissue.

CART was detected in perfusion fixed frozen sections of mouse brain tissue using Goat Anti-Human/Mouse/Rat CART Antigen Affinity-purified Polyclonal Antibody (Catalog # AF163) at 5 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to neurons. Staining was performed using our Fluorescent IHC Staining of Frozen Tissue Sections.
Detection of CART (aa 28-116) by Western Blot

Detection of CART (aa 28-116) by Western Blot

Characterization of CART peptide (CARTp) antibodies. A PCR amplification curves of CART mRNA extracted from donor cadaver DMN and celiac ganglion tissues. CARTp mRNA was detected in the DMN, but not in the celiac ganglion. B Representative capillary westerns showing CARTp immunoreactivity for three commercially available CART antibodies. Synthetic CARTp was denatured and loaded as the sample. C Representative capillary westerns showing the reactivity of the mouse anti-CARTp antibody against different amounts of synthetic CARTp and scrambled CARTp. D Representative images of the paraventricular nucleus (PVN, a site known to contain CART-positive axons) showing CARTp immunoreactivity in tissue sections incubated with either the mouse anti-CART antibody (CART Ab), the mouse anti-CART antibody after overnight neutralization with 30 molar excess CART peptide (CART Ab + CARTp), or the mouse anti-CART antibody after overnight neutralization with 30 molar excess scrambled CART peptide (CART Ab + Scr pep) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37403174), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of CART (aa 28-116) by Immunohistochemistry

Detection of CART (aa 28-116) by Immunohistochemistry

CART peptide (CARTp) is expressed in neurons in the caudal DMN. A Drawing of a coronal section of the human brain stem demonstrating the relative position of the DMN (rostral aspect in red) compared to the NTS and the 4th ventricle. B Representative images showing that CARTp immunoreactivity can be detected in neurons in the caudal, but not rostral, aspect of the DMN in humans. The insets show that both CARTp-positive cell somas (arrows) and fibers (arrowheads) can be seen in caudal DMN. C Representative mouse brain images showing CARTp immunoreactivity in neurons resident to the caudal, but not rostral, aspect of the DMN. D Intracerebroventricular administration of CART siRNA to mice significantly reduced CARTp immunoreactivity in the caudal DMN compared to control (non-targeting) siRNA. E Representative double-label immunohistochemistry showing co-localization of CARTp and ChAT immunoreactivities in the caudal DMN neurons in mice. Double immunostained cells appear yellow in the merged image Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37403174), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for CART Antibody

Application
Recommended Usage

Immunohistochemistry

1-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human brain (Hypothalamus) tissue and perfusion fixed frozen sections of mouse brain tissue

Western Blot

0.1 µg/mL
Sample: Recombinant Human CART

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CART

CART, also known as cocaine and amphetamine-regulated transcript, is a 10 kDa secreted polypeptide produced by neuronal cell types in a variety of locations. In human, the molecule is synthesized as a 116 amino acid (aa) precursor with a 27 aa signal sequence an 89 aa mature segment. There are six C-terminal cysteines that form three intrachain disulfide bonds, and the potential exists for both sulfation and phosphorylation in the N-terminal region. There are numerous monobasic and dibasic sites for enzyme cleavage, and multiple, presumably proteolytically-generated short forms are known to exist. These include a 5K, 48 aa peptide (CART 42‑89), and a 4.3K, 40 aa peptide (CART 49-89). Human CART is 98% aa identical to both moyuse and rat CART (1‑89). Although CART suppresses feeding and increases pain tolerance, there would appear to be differences between the various forms of CART on other functions. In addition, anatomical location is associated with differing lengths of CART. For example, 89 aa and 79 aa forms are found in the adrenal, while 48 aa and 44 aa forms are found in the hypothalamus.

References

  1. Douglass, J. & S. Daoud (1996) Gene 169:241.
  2. Douglass, J. et al. (1995) J. Neurosci. 15:247.
  3. Adams, L.D. et al. (1999) Brain Res. 848:137.
  4. Thim, L. et al. (1999) Proc. Natl. Acad. Sci. USA 96:2722.
  5. Bannon, A.W. et al. (2001) J. Pharmacol. Exp. Ther. 299:1021.
  6. Kuhar, M.J. & L.L. Yoho (1999) Synapse 33:163.
  7. Thim, L. et al. (1998) FEBS Lett. 428:263.

Long Name

Cocaine- and Amphetamine-regulated Transcript Protein

Alternate Names

CARTPT

Entrez Gene IDs

9607 (Human)

Gene Symbol

CARTPT

UniProt

Q16568

Additional CART Products

Product Documents for CART Antibody

Certificate of Analysis

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Product Specific Notices for CART Antibody

For research use only

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