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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 yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)DYTWNKDNWTFSP, corresponding to amino acid residues 72 - 84 of human GPR65 (Accession Q8IYL9). 1st extracellular 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.
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).
Standard quality control of each lot Western blot analysis.
Applications: if, ifc, ih, wb
May also work in: ic*, ip*, lci*
Western blot
  • 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:
    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 GPR65/TDAG8 (extracellular) Blocking Peptide (#BLP-GR043).
  • Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain membranes:
    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 GPR65/TDAG8 (extracellular) Blocking Peptide (#BLP-GR043).
Immunohistochemistry
  • Expression of GPR65 in rat fornix
    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).
Indirect flow cytometry
  • Cell surface detection of GPR65 in live intact human Jurkat T-cell leukemia cells:
    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
  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, 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: 23/08/2020

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

Applications

Specifications

Scientific Background

Citations

Citations
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