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Anti-ENPP1 (extracellular) Antibody

Ectonucleotide Pyrophosphatase/ Phosphodiesterase Family Member 1, Plasma-Cell Membrane Glycoprotein PC-1, CD203a, Membrane Component Chromosome 6 Surface Marker 1, M6S1

Cat #: ANT-069
Alternative Name Ectonucleotide Pyrophosphatase/ Phosphodiesterase Family Member 1, Plasma-Cell Membrane Glycoprotein PC-1, CD203a, Membrane Component Chromosome 6 Surface Marker 1, M6S1
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide CLYQDFRIPLSPVHK, corresponding to amino acid residues 711 - 725 of human ENPP1 (Accession P22413). Extracellular, C-term.
Accession (Uniprot) Number P22413
Gene ID 5167
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse – 14 out of 15 amino acid residues identical; Rat- 13 out of 15 amino acid residues identical.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Preadsorption Control 1 µg peptide per 1 µg antibody
ENPP1 (extracellular) Blocking Peptide (#BLP-NT069)
Standard quality control of each lot Western blot analysis.
Applications: ifc, ih, lci, wb
May also work in: ic*, ip*
Western blot
  • Western blot analysis of mouse brain lysate (lanes 1 and 3) and rat brain lysate (lanes 2 and 4):
    Western blot analysis of mouse brain lysate (lanes 1 and 3) and rat brain lysate (lanes 2 and 4):
    1-2. Anti-ENPP1 (extracellular) Antibody (#ANT-069), (1:400).
    3-4. Anti-ENPP1 (extracellular) Antibody, preincubated with ENPP1 Blocking Peptide (BLP-NT069).
  • Western blot analysis of human THP-1 monocytic leukemia cell line lysate (lanes 1 and 4), human HL-60 promyelocytic leukemia cell line lysate (lanes 2 and 5) and human MDA-MB-231 breast adenocarcinoma cell line lysate (lanes 3 and 6):
    Western blot analysis of human THP-1 monocytic leukemia cell line lysate (lanes 1 and 4), human HL-60 promyelocytic leukemia cell line lysate (lanes 2 and 5) and human MDA-MB-231 breast adenocarcinoma cell line lysate (lanes 3 and 6):
    1-3. Anti-ENPP1 (extracellular) Antibody (#ANT-069), (1:200).
    4-6. Anti-ENPP1 (extracellular) Antibody, preincubated with ENPP1 Blocking Peptide (BLP-NT069).
Immunohistochemistry
  • Expression of ENPP1 in rat spinal cord.
    Expression of ENPP1 in rat spinal cord.
    Immunohistochemical staining of perfusion-fixed frozen rat spinal cord sections with Anti-ENPP1 (extracellular) Antibody (#ANT-069), (1:1000), followed by goat anti-rabbit-AlexaFluor-488. A. Staining in the dorsal horn region, showed ENPP1 immunoreactivity (green) neuronal profiles (vertical arrows) and glial profiles (horizontal arrows). B. Pre-incubation of the antibody with ENPP1 Blocking Peptide (BLP-NT069), suppressed staining. Cell nuclei are stained with DAPI (blue).
Indirect flow cytometry
  • Cell surface detection of ENPP1 by indirect flow cytometry in live intact human THP-1 monocytic leukemia cell line:
    Cell surface detection of ENPP1 by indirect flow cytometry in live intact human THP-1 monocytic leukemia cell line:
    ___ Cells.
    ___ Cells + goat-anti-rabbit-APC.
    ___ Cells + Anti-ENPP1 (extracellular) Antibody (#ANT-069) (2.5μg) + goat-anti-rabbit-APC.
References
  1. Borza, R. et al. (2022) J. Biol. Chem. 298, 101526.
  2. Onyedibe, K.I. et al. (2019) Molecules. 24, 4192.
  3. Roberts, F. et al. (2019) Trends Bio. Sci. 44, 616.
  4. Carozza, J.A. et al. (2022) PNAS. 119, e2119189119.
  5. Kato, K. et al. (2018) Nat. Commun. 9, 4424.
  6. Zhou, Y. et al. (2020) Immunity. 52, 357.
  7. Ritchie, C. et al. (2019) Mol. Cell. 75, 372.
  8. Zhou, C. et al. (2020) Immunity. 52, 767.
  9. Carozza, J.A. et al. (2020) Nat. Cancer. 1, 184.
Scientific background

Ectonucleotide Pyrophosphatase/Phosphodiesterase 1 (ENPP1), also known as Plasma-Cell Membrane Glycoprotein (PC-1) and CD203a, is a member the ecto-enzyme ENPP family that includes seven members (ENPP1-ENPP7)1.

ENPP1 is involved in purinergic signaling as it catalyzes the hydrolysis of ATP or GTP to AMP or GMP, while generating inorganic pyrophosphates (PPi)1,2. Inorganic pyrophosphates inhibit bone and cartilage mineralization, hence making ENPP1 a key regulator of bone and cartilage development and remodeling 2,3. Indeed, several hereditary mineralization or calcification-related disorders, have been linked to loss-of-function mutations of ENPP1, such as  general arterial calcification of infancy (GACI) or Cole disease 3.

Recently, an intriguing new role of ENPP1 as a central regulator of immunity and tumor proliferation came to light. It was shown that ENPP1 is the critical enzyme that degrades the stimulator of interferon genes (STING) ligand, cyclic GMP–AMP (cGAMP)4,5. cGAMP was first described as an intracellular second messenger, synthesized by the enzyme cyclic-GMP-AMP synthase (cGAS) in response to self and pathogenic double-stranded DNA in the cytosol. Intracellular signaling of cGAMP is carried through the STING pathway which then directs several antiviral and anticancer cellular response mechanisms 4,5.

cGAMP can also be released to the extracellular space where it can be transported into cells by receptors like MERTK 6, P2X7 6, SLC19A1 7 and LRRC8A 8. ENPP1 is the only known ecto-enzyme that can inactivate extracellular cGAMP to GMP and AMP, and thus functions as an inhibitor of the anti-tumor response elicited by extracellular cGAMP 4,9.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 08/01/2023

Anti-ENPP1 (extracellular) Antibody (#ANT-069) is a highly specific antibody directed against an extracellular epitope of the human protein. The antibody can be used in western blot, immunohistochemistry and flow cytometry applications. It has been designed to recognize ENPP1 from mouse, rat and human samples.

For research purposes only, not for human use
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