Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry

Cited:

Western Blot, ELISA Capture

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
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Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human GPR64
Leu38-Asn64, Glu68-Thr553
Accession # NP_001073328

Specificity

Detects human, mouse, and rat GPR64 in direct ELISA and Western blots.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat GPR64 Antibody

Detection of Rat and Mouse GPR64 antibody by Western Blot.

Detection of Rat and Mouse GPR64 by Western Blot.

Western blot shows lysates of rat epididymis tissue and mouse epididymis tissue. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human/Mouse/Rat GPR64 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7977) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for GPR64 at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

GPR64 antibody in Human Epididymus by Immunohistochemistry (IHC-P).

GPR64 in Human Epididymus.

GPR64 was detected in immersion fixed paraffin-embedded sections of human epididymus using Sheep Anti-Human/Mouse/Rat GPR64 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7977) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membranes of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of GPR64 antibody in HEK293 Human Cell Line Transfected with Human GPR64 and eGFP antibody by Flow Cytometry.

Detection of GPR64 in HEK293 Human Cell Line Transfected with Human GPR64 and eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with (A) Human GPR64 or (B) Human GPR75 and eGFP was stained with Sheep Anti-Human GPR64 Affinity-Purified Polyclonal Antibody (Catalog # AF7977) followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). Quadrant markers were set based on control antibody staining (Catalog # 5-001-A). View our protocol for Staining Membrane-associated Proteins.

Detection of GPR64 by Western Blot

Detection of GPR64 by Western Blot

The expression of ADGRG2, CFTR, Gs, Gq, beta -arrestin-1, beta -arrestin-2 in efferent ductules, brain and liver tissue of WT and Adgrg2-/Y mice.(A) Western blot analysis of ADGRG2, CFTR, Gs, Gq, beta -arrestin-1, beta -arrestin-2 expression in efferent ductules, brain and liver tissue of WT and Adgrg2-/Y mice. A representative western blot from at least three independent experiments was shown. (CFTR antibody:20738–1-AP, Proteintech). (B) Bar graph representation and statistical analyses of (A). All blots were normalized to GAPDH. n.s., no significant difference; Adgrg2-/Y mice compared with WT mice in the same tissue. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29393851), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GPR64 by Western Blot

Detection of GPR64 by Western Blot

The complex formation between ADGRG2, beta -arrestin-1 and CFTR in HEK293 cells.(A) HEK293 cells were transfected with equal amount plasmids encoding ADGRG2, CFTR, beta -arrestin-1 or beta -arrestin-2 plasmids. The Flag- ADGRG2 were pulled down by M2-Flag beads and the associated CFTR, beta -arrestins were detected by western blot. Representative images from at least three independent experiments are shown (CFTR antibody:20738–1-AP, Proteintech). (B) Bar graph representation and statistical analyses of (A).***p<0.001, IP protein of ADGRG2 overexpressed cells were compared with control plasmids transfected cells respectively. n.s., no significant difference. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29393851), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GPR64 by Western Blot

Detection of GPR64 by Western Blot

The ADGRG2 protein knockout strategy, PCR strategy and western blot results of Adgrg2-/Y and Gnaq+/- mice.(A) Schematic representation of the ADGRG2 knockout strategy for the Adgrg2-/Y mice. 2 bp nucleotides were removed and 10 bp nucleotides were inserted in the first exon of the ADGRG2 gene in the ADGRG2 mutant mice by the CRISP-CAS9 approach. In the ADGRG2-deficient mice, the translation of ADGRG2 was terminated at the 7th amio acid after the signal peptide. (B) Schematic representation for the primers used in the genotyping of the ADGRG2 mutant mice or their wild-type littermates. (C) Schematic description of the PCR strategy and expected results for genotyping. The genotyping of mice was determined by PCR and visualized by bromide staining of agarose Gels. (D) Western blot analysis of ADGRG2 expression in efferent duct tissue of WT and Adgrg2-/Y mice. All blots were normalized to GAPDH. (E) Bar graph representation and statistical analyses of (D). At least three independent experiments were carried out. (F) Western blot analysis of Gq expression in efferent duct tissue of WT and Gnaq+/- mice. All blots were normalized to GAPDH. (G) Bar graph representation and statistical analyses of (F). At least three independent experiments were carried out. (E,G) *p<0.05, **p<0.01, ***p<0.001, Adgrg2-/Y mice or Gnaq+/- mice were compared with WT mice. n.s., no significant difference. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29393851), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat GPR64 Antibody

Application
Recommended Usage

Flow Cytometry

0.25 µg/106 cells
Sample: HEK293 Human Cell Line Transfected with Human GPR64 and eGFP

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human epididymus

Western Blot

2 µg/mL
Sample: Rat epididymis tissue and mouse epididymis tissue

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: GPR64

GPR64 (G-Protein coupled Receptor 64; also HE6) is a 110 kDa (predicted) member of the B class of GCPRs. Within this class GPR64 belongs to a Large N-termini family-B 7-transmembrane (LNB-7TM) subclass of receptors (also known as adhesion-GCPRs). GPR64 has restricted expression, being found in stereocilia cell membranes of epididymal caput epithelial cells and, to a limited extent, on osteoblasts. The function of GPR64 is somewhat unclear, but in the epididymis, it may be involved in fluid transport. Mature human GPR64 is a 980 amino acid (aa) 7-TM glycoprotein (SwissProt Q8IZP9). It contains an extended extracellular N-teminus (aa 38-627), seven consecutive TM segments (aa 628-878) and a C-terminal cytoplasmic tail (aa 879-1010). The extended extracellular region possesses a juxtamembrane GPS domain (aa 567-618) that serves as a proteolytic cleavage site. Enzymatic activity here generates a 180 kDa soluble form that stays associated with a 40 kDa membrane-embedded fragment. Notably, isolation of the membrane fragment gives rise to oligomers that run at > 250 kDa in SDS-PAGE. There are multiple splice variants. The one used for immunization to generate this antibody contains a deletion of aa 88-101 (RefSeq NP_001073328). Four other splice forms show single block deletions of aa 65-67, 51-66, 52-75, and 906-956, respectively. Three others possesses aa substitutions; a 20 aa block for aa 52-101, and a common 12 aa block that can substitute for either aa 68-101 or aa 52-101. Over aa 38-64 and 68-553 of RefSeq NP_001073328, human and mouse GPR64 share 69% aa sequence identity.

Long Name

G Protein-coupled Receptor 64

Alternate Names

EDDM6, HE6, TM7LN2

Entrez Gene IDs

10149 (Human); 237175 (Mouse); 266735 (Rat)

Gene Symbol

ADGRG2

UniProt

Additional GPR64 Products

Product Documents for Human/Mouse/Rat GPR64 Antibody

Certificate of Analysis

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Product Specific Notices for Human/Mouse/Rat GPR64 Antibody

For research use only

Citations for Human/Mouse/Rat GPR64 Antibody

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Protocols

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