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Anti-Adenosine A2B Receptor (extracellular) Antibody

ADORA2b, A2BAR, Adenosine A2bR

Cat #: AAR-003
Alternative Name ADORA2b, A2BAR, Adenosine A2bR
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide KDSATNN*STEPWDGTTNESC, corresponding to amino acid residues 147-166 of human A2BAR with replacement of cysteine 154 (C154) with serine (*S) (Accession P29275). 2nd extracellular loop.
Accession (Uniprot) Number P29275
Gene ID 136
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat - 14/19 amino acid residues identical; mouse - 13/19 amino acid residues identical.
RRID AB_2039709.
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 0.75 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).
Standard quality control of each lot Western blot analysis.
Applications: ic, if, ifc, ih, lci, wb
May also work in: ip*
Western blot
  • Western blot analysis of HL-60 (lanes 1 and 2) cell line, rat brain (lanes 3 and 4) and mouse brain (lanes 5 and 6) lysates:
    Western blot analysis of HL-60 (lanes 1 and 2) cell line, rat brain (lanes 3 and 4) and mouse brain (lanes 5 and 6) lysates:
    1,3,5. Anti-Adenosine A2B Receptor (extracellular) Antibody (#AAR-003), (1:200).
    2,4,6. Anti-Adenosine A2B Receptor (extracellular) Antibody, preincubated with Adenosine A2B Receptor (extracellular) Blocking Peptide (#BLP-AR003).
Immunohistochemistry
  • Expression of Adenosine A2B Receptor in rat lung
    Expression of Adenosine A2B Receptor in rat lung
    Immunohistochemical staining of rat lung paraffin embedded sections using Anti-Adenosine A2B Receptor (extracellular) Antibody (#AAR-003), (1:50). Staining is present in the respiratory epithelium of the bronchiole (Br) as well as in the pneumonocytes of the alveolar wall (alveoli, (A). Color reaction was obtained with SuperPicture HRP-conjugated polymer (Zymed) followed by DAB. Hematoxilin is used as the counterstain.
Indirect flow cytometry
  • Rat enterochromaffin (EC) cells (1:500), (Damen, R. et al. (2013) PLoS ONE 8, e62607.).
  • Cell surface detection of Adenosine A2B Receptor by indirect flow cytometry in live intact human THP-1 monocytic leukemia cells:
    Cell surface detection of Adenosine A2B Receptor by indirect flow cytometry in live intact human THP-1 monocytic leukemia cells:
    ___ Cells.
    ___ Cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-Adenosine A2B Receptor (extracellular) Antibody (#AAR-003), (2.5μg) + goat-anti-rabbit-FITC.
Live cell imaging / Immunocytochemistry
  • Expression of Adenosine A2B Receptor in rat PC12 cells
    Expression of Adenosine A2B Receptor in rat PC12 cells
    Cell surface detection of Adenosine A2B Receptor in intact living rat pheochromocytoma PC12 cells. A. Extracellular staining of cells using Anti-Adenosine A2B Receptor (extracellular) Antibody (#AAR-003), (1:100) followed by goat anti-rabbit-AlexaFluor-594 secondary antibody. B. Merge of A and live view of the cells.
References
  1. Peeters, M.C. et al. (2011) FASEB J. 25, 632.
  2. Sun, Y. and Huang, P. (2016) Front. Chem. 4, 37.
Scientific background

Extracellular adenosine is a ubiquitous signaling molecule that modulates a wide array of biological processes. There are currently four known sub-types of adenosine receptors: A1, A2A, A2B and A3.

The adenosine A2B receptor is a typical class A, G protein-coupled receptor with seven transmembrane segments, an extracellular N-terminus and an intracellular C-terminus. Adenosine A2BR causes an increase in intracellular cAMP levels by coupling mainly to Gs proteins, resulting in the activation of adenylyl cyclase1 and subsequent activation of PKA and other cAMP effectors such as Epac. In addition, the A2BR couples to Gq and activates the PLC pathway that mediates several crucial functions of A2BR. A2BR also couples to the MAPK and arachidonic acid signaling pathways and regulates membrane ion channels probably through G-protein βγ subunits.

Multiple molecules that bind to A2BR have been identified in an effort to understand the receptor’s function and regulation. NFκB1/p105 is a member of the NFκB family of proteins that perform regulatory functions in diverse biological processes such as inflammation and cell survival and differentiation, as well as in various diseases. Interestingly, Although A2BR activation by adenosine produces pro-inflammatory effects the receptor also binds to specific sites on p105, prevents the poly-ubiquitination and degradation of the p105 protein and inhibits NFκB activation thus reducing inflammation.

A2BR is highly expressed in various types of tumor cells or tissues and promotes tumor-cell proliferation. A2BR was found to be overexpressed in colorectal carcinoma cells and tissues, and inhibition of A2BR blocks the proliferation of colon cancer cells. A2BR has been implicated in the pathophysiology of vascular diseases, renal disease and diabetes2.

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: 05/05/2021

Anti-Adenosine A2B Receptor (extracellular) Antibody (#AAR-003) is a highly specific antibody directed against an extracellular epitope of human adenosine A2B receptor (A2BAR). The antibody can be used in western blot, immunohistochemistry, live cell imaging and indirect flow cytometry applications. It has been designed to recognize A2BAR from human, rat and mouse samples.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
Western blot citations
  1. Human ovarian cancer A1780 and HEY cell lysates.
    Sureechatchaiyan, P. et al. (2018) Purinergic Signal. 14, 395.
  2. Human skin lysate.
    Andres, R.M. et al. (2017) J. Invest. Dermatol. 137, 123.
  3. Rat lung lysate.
    Densmore, J.C. et al. (2017) Exp. Lung Res. 43, 38.
  4. Rat retina cell lysate (1:1000).
    Vindeirinho, J. et al. (2013) PLoS ONE 8, e67499.
Immunohistochemistry citations
  1. Rat pancreas sections (1:800).
    Hayashi, M. et al. (2016) Eur. J. Physiol. 468, 1171.
Immunocytochemistry citations
  1. Human cholangiocyte cells.
    Lavoie, E.G. et al. (2017) Gene Expres. 17, 327.
  2. Rat neurospheres.
    Benito-Munoz, M. et al. (2016) Glia 64, 1465.
Indirect flow cytometry citations
  1. Mouse lymph node dendritic cells.
    Gardner, J.K. et al. (2018) Front. Med. (Lausanne) 5, 337.
More product citations
  1. Damen, R. et al. (2013) PLoS ONE 8, e62607.
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