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Anti-GPR109A/HCAR2 (extracellular) Antibody

Hydroxycarboxylic Acid Receptor 2, Niacin Receptor 1, G-Protein Coupled Receptor HM74A, HM74A, Nicotinic acid receptor, PUMAG

Cat #: AHR-012
Alternative Name Hydroxycarboxylic Acid Receptor 2, Niacin Receptor 1, G-Protein Coupled Receptor HM74A, HM74A, Nicotinic acid receptor, PUMAG
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)DNYVHNWDWRFGG, corresponding to amino acid residues 82 - 94 of mouse HCAR2 (Accession Q9EP66). Extracellular, 1st loop.
Accession (Uniprot) Number Q9EP66
Gene ID 80885
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat – 11 out of 13 amino acid residues identical; Human - 9 out of 13 amino acid residues identical.  
Note: it is not expected to recognize the highly homologous human protein HCAR3 (Accession P49019). HCAR3 has no mouse or rat orthologs.
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
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 rat spleen lysates (lanes 1 and 3) and rat colon membranes (lanes 2 and 4):
    Western blot analysis of rat spleen lysates (lanes 1 and 3) and rat colon membranes (lanes 2 and 4):
    1-2. Anti-GPR109A/HCAR2 (extracellular) Antibody (#AHR-012), (1:200).
    3-4. Anti-GPR109A/HCAR2 (extracellular) Antibody, preincubated with GPR109A/HCAR2 (extracellular) Blocking Peptide (BLP-HR012).
  • Western blot analysis of mouse colon lysates (lanes 1 and 3) and mouse lung lysates (lanes 2 and 4):
    Western blot analysis of mouse colon lysates (lanes 1 and 3) and mouse lung lysates (lanes 2 and 4):
    1-2. Anti-GPR109A/HCAR2 (extracellular) Antibody (#AHR-012), (1:200).
    3-4. Anti-GPR109A/HCAR2 (extracellular) Antibody, preincubated with GPR109A/HCAR2 (extracellular) Blocking Peptide (BLP-HR012).
  • Western blot analysis of human THP-1 monocytic leukemia cell line lysates (lanes 1 and 3) and human HL-60 promyelocytic leukemia cell line lysates (lanes 2 and 4):
    Western blot analysis of human THP-1 monocytic leukemia cell line lysates (lanes 1 and 3) and human HL-60 promyelocytic leukemia cell line lysates (lanes 2 and 4):
    1-2. Anti-GPR109A/HCAR2 (extracellular) Antibody (#AHR-012), (1:200).
    3-4. Anti-GPR109A/HCAR2 (extracellular) Antibody, preincubated with GPR109A/HCAR2 (extracellular) Blocking Peptide (BLP-HR012).
Immunohistochemistry
  • Expression of GPR109A in rat cingulate cortex.
    Expression of GPR109A in rat cingulate cortex.
    Immunohistochemical staining of perfusion-fixed frozen rat brain sections with Anti-GPR109A/HCAR2 (extracellular) Antibody (#AHR-012), (1:200), followed by goat anti-rabbit-AlexaFluor-488. A. GPR109A immunoreactivity (green) appears in outlines of cortical neurons (arrows). B. Pre-incubation of the antibody with GPR109A/HCAR2 (extracellular) Blocking Peptide (BLP-HR012), suppressed staining. Cell nuclei are stained with DAPI (blue).
Indirect flow cytometry
  • Cell surface detection of GPR109A by indirect flow cytometry in live intact mouse J774 macrophage cell line:
    Cell surface detection of GPR109A by indirect flow cytometry in live intact mouse J774 macrophage cell line:
    ___ Cells.
    ___ Cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti- GPR109A/HCAR2 (extracellular) Antibody (#AHR-012), (2.5μg) + goat-anti-rabbit-FITC.
References
  1. Ahmed K. (2011) Front. Endocrinol. 25, 51.
  2. Graff E. et al (2016) Metabolism. 65, 102-113.
  3. Offermanns S., (2017) Trends Endocrinol Metab. 28, 227-236.
  4. Offermanns S.,(2014) Annu. Rev. Pharmacol. Toxicol. 54, 407–34.
Scientific background

Hydroxy-carboxylic acid receptors (HCA1-3) are a family of G-protein-coupled receptors that regulate lipolysis. HCAR2 (GPR109A) was identified as the receptor for nicotinic acid (niacin; vitamin B3) and beta-hyroxybutyrate (β-OHB), a ketone body synthesized by the liver under negative energy balance1.

HCAR2 is mainly expressed on adipocytes and immune cells and in lower levels in retinal pigmented and colonic epithelial cells, microglia and normal mammary tissue. High levels of HCAR2 can be found in immune cells such as macrophages in response to the presence of interferon-γ, TNF-α and hypoxia2.

HCAR2 activity in adipocytes promotes inhibition of adenylate cyclase and decrease cAMP levels. HCAR2 activity leads to a rapid decrease in lipolysis and reduction in the release of fatty acids from the adipocytes3.

Activation of HCAR2 has also been associated with anti-inflammatory effects in different disease states such as sepsis, diabetes and obesity. Interaction of HCAR2 with niacin and β-OHB under this conditions lead to variety of signaling events involving inflammatory downstream targets4.

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: 08/06/2021

Anti-GPR109A/HCAR2 (extracellular) Antibody (#AHR-012) is a highly specific antibody directed against an extracellular epitope of the mouse protein. The antibody can be used in western blot, immunohistochemistry and flow cytometry applications. It has been designed to recognize GPR109A from rat, mouse and human samples.

For research purposes only, not for human use
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