Anti-GPR65 (TDAG8) (extracellular) Antibody

G-protein coupled receptor 65, T-cell death-associated gene 8 protein, Psychosine receptor
    Cat #: AGR-043
    Alternative Name G-protein coupled receptor 65, T-cell death-associated gene 8 protein, Psychosine receptor
  • Lyophilized Powder
  • Antigen Incl.
  • Type: Polyclonal
    Host: Rabbit
    Reactivity: h, m, r
      • Peptide (C)DYTWNKDNWTFSP, corresponding to amino acid residues 72 - 84 of human GPR65 (Accession Q8IYL9). 1st extracellular loop.
        Anti-GPR65 (TDAG8) (extracellular) Antibody
    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.
    RRID AB_2756633.
    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).
    Negative 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.
    Negative control antigen reconstitution 100 µl double distilled water (DDW).
    Negative control antigen storage after reconstitution -20°C.
    Preadsorption Control 1 µg peptide per 1 µg antibody.
    Standard quality control of each lot Western blot analysis.
    Applications: if, ifc, ih, wb
    May also work in: ic*, ip*, lci*
      • Anti-GPR65 (TDAG8) (extracellular) Antibody
        Western blot analysis of human Jurkat T-cell leukemia (lanes 1 and 4), human THP-1 monocytic leukemia (lanes 2 and 5) and human HL-60 promyelocytic leukemia (lanes 3 and 6) cell line lysates:
        1-3. Anti-GPR65 (TDAG8) (extracellular) Antibody (#AGR-043), (1:200).
        4-6. Anti-GPR65 (TDAG8) (extracellular) Antibody, preincubated with the negative control antigen.
        Anti-GPR65 (TDAG8) (extracellular) Antibody
        Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain membranes:
        1,2. Anti-GPR65 (TDAG8) (extracellular) Antibody (#AGR-043), (1:200).
        3,4. Anti-GPR65 (TDAG8) (extracellular) Antibody, preincubated with the negative control antigen.
      • Anti-GPR65 (TDAG8) (extracellular) Antibody
        Expression of GPR65 in rat fornix
        Immunohistochemical staining of immersion-fixed, free floating rat frozen brain sections using Anti-GPR65 (TDAG8) (extracellular) Antibody (#AGR-043), (1:1200), followed by goat-anti-rabbit-AlexaFluor-488. GPR65 staining (green) appears in glial processes (vertical arrows) and in the side of fornix (FX) facing the ventricle (V, horizontal arrow). Cell nuclei are stained with DAPI (blue).
      • Anti-GPR65 (TDAG8) (extracellular) Antibody
        Cell surface detection of GPR65 in live intact human Jurkat T-cell leukemia cells:
        ___ Cells.
        ___ Cells + goat-anti-rabbit-FITC.
        ___ Cells + Anti-GPR65 (TDAG8) (extracellular) Antibody (#AGR-043), 5 µg + goat-anti-rabbit-FITC.
    References
      • 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, 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: 24/01/2020

    Anti-GPR65 (TDAG8) (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 GPR65 from rat, mouse, and human samples.

    For research purposes only, not for human use