Anti-GPR65 (extracellular)-FITC Antibody

G Protein-Coupled Receptor 65, T-Cell Death-Associated Gene 8 Protein, TDAG8, Psychosine Receptor
    Cat #: AGR-043-F
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    Peptide (C)DYTWNKDNWTFSP, corresponding to amino acid residues 72 - 84 of human GPR65 (Accession Q8IYL9). Extracellular, 1st loop.
    Accession (Uniprot) Number Q8IYL9
    Gene ID 8477
    Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
    Homology Mouse – 12/13 amino acid residues identical; rat - 11/13 amino acid residues identical.
    Purity Affinity purified on immobilized antigen.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
    Label Fluorescein isothiocyanate (FITC).
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 µl double distilled water (DDW).
    Antibody concentration after reconstitution 1 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × g 5 min).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 µl double distilled water (DDW).
    Control antigen storage after reconstitution -20ºC.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis (unlabeled antibody, #AGR-043), and direct flow cytometry (labeled antibody).
    Applications: fc
    May also work in: ic, lci
    Direct flow cytometry
    Direct flow cytometry of live intact human Jurkat T-cell leukemia cells:
    ___ Cells.
    ___ Cells + rabbit IgG isotype control-FITC.
    ___ Cells + Anti-GPR65 (extracellular)-FITC Antibody (#AGR-043-F), (5 µg).
    Direct flow cytometry of live intact mouse M1 myeloid leukemia cells:
    ___ Cells.
    ___ Cells + rabbit IgG isotype control-FITC.
    ___ Cells + Anti-GPR65 (extracellular)-FITC Antibody (#AGR-043-F), (5 µg).
    References
    1. Justus, C.R. et al. (2013) Front. Physiol. 4, 354.
    2. Justus, C.R. et al. (2017) J. Transl. Med. 15, 204.
    Scientific background

    GPR65, also called TDAG8 (T-cell death associated gene 8 protein), is a pH-sensing G-protein coupled receptor (GPCR) activated by acidic extracellular pH through the protonation of several histidine residues in the receptor’s sequence. GPR65 is involved in cancer cell metastasis and proliferation, immune cell function, inflammation, and blood vessel formation1.

    Human GPR65 gene has been mapped to a location that is associated with T cell lymphoma and leukemia abnormalities.

    GPR65 is primarily expressed in immune cells and leukocyte-rich tissues such as circulating peripheral leukocytes, spleen, thymus, and tonsils. Expression is also detected pain relevant loci such as the dorsal root ganglia neurons and particularly small diameter neurons responsible for nociception1,2.

    GPR65 has the ability to affect tumor development and growth. Overexpression of GPR65 in Lewis lung carcinoma cells increases tumor growth in mice and may facilitate resistance to acidosis-mediated cell death in vitro through protein kinase A (PKA) and ERK related pathways1.

    Knockdown of GPR65 in NCI-H460 human non-small cell lung cancer cells decreases cell survival in acidic condition. Another study shows that activation of GPR65 by acidosis can promote evasion of cell apoptosis under glutamine starvation and its overexpression has been reported to transform immortalized mammary epithelial cells1.

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, 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: 07/08/2018

    Anti-GPR65 (extracellular) Antibody (#AGR-043) is a highly specific antibody directed against an epitope of the human protein. The antibody can be used in western blot, immunohistochemistry, and live cell flow cytometry. It has been designed to recognize TDAG8 from rat, mouse, and human samples.

    Anti-GPR65 (extracellular)-FITC Antibody (#AGR-043-F) is directly conjugated to fluorescein isothiocyanate (FITC). This labeled antibody can be used in immunofluorescent applications such as direct live cell flow cytometry.

    For research purposes only, not for human use