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Anti-GPRC5B Antibody

G-Protein Coupled Receptor Family C Group 5 Member B, Retinoic Acid-Induced Gene 2 Protein, RAIG2

Cat #: AGR-072
Alternative Name G-Protein Coupled Receptor Family C Group 5 Member B, Retinoic Acid-Induced Gene 2 Protein, RAIG2
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)ENPPNYFDTSQPRMRE, corresponding to amino acid residues 302 - 317 of mouse GPRC5B (Accession Q923Z0). Intracellular, C-terminus
Accession (Uniprot) Number Q923Z0
Gene ID 64297
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat – identical; Human - 15 out of 16 amino acid residues identical.
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
GPRC5B Blocking Peptide (#BLP-GR072)
Standard quality control of each lot Western blot analysis.
Applications: ih, wb
May also work in: ic*, ip*
Western blot
  • Western blot analysis of rat brain lysates (lanes 1 and 3) and mouse brain lysates (lanes 2 and 4):
    Western blot analysis of rat brain lysates (lanes 1 and 3) and mouse brain lysates (lanes 2 and 4):
    1-2. Anti-GPRC5B Antibody (#AGR-072), (1:800).
    3-4. Anti-GPRC5B Antibody, preincubated with GPRC5B Blocking Peptide (BLP-GR072).
  • Western blot analysis of human K562 myelogenous leukemia cell line lysate (lanes 1 and 3) and human NCI-H526 lung carcinoma cell line lysates (lanes 2 and 4):
    Western blot analysis of human K562 myelogenous leukemia cell line lysate (lanes 1 and 3) and human NCI-H526 lung carcinoma cell line lysates (lanes 2 and 4):
    1-2. Anti-GPRC5B Antibody (#AGR-072), (1:200).
    3-4. Anti-GPRC5B Antibody, preincubated with GPRC5B Blocking Peptide (BLP-GR072).
Immunohistochemistry
  • Expression of GPRC5B in mouse deep cerebellar nuclei.
    Expression of GPRC5B in mouse deep cerebellar nuclei.
    Immunohistochemical staining of perfusion-fixed frozen mouse brain sections using Anti-GPRC5B Antibody (#AGR-072), (1:200), followed by goat anti-rabbit-AlexaFluor-488. A. GPRC5B immunoreactivity (green) appears in neuron ourlines (arrows). B. Pre-incubation of the antibody with GPRC5B Blocking Peptide (BLP-GR072), suppressed staining. Cell nuclei are stained with DAPI (blue).
References
  1. Robbins, M J et al. (2000) Genomics, 67, 1.
  2. Alonso-Gardón, Marta et al. (2021) Human molecular genetics. 30, 17.
  3. Hirabayashi, Yoshio, and Yeon-Jeong Kim. (2020) Journal of biochemistry. 167, 6.
  4. Kim, Yeon-Jeong et al. (2012) Science signaling. 5, 251.
Scientific background

GPRC5B is a member of the large G-protein-coupled receptor (GPCR) superfamily of receptors that share a common structure of seven membrane-spanning domains, an extracellular N-terminal domain, an intracellular C-terminal domain, and many conserved residues[1].

The ligand-binding domain of GPCRs is variable and the GPCRs have been divided into three classes based on the ligands that stimulate them, as well by key sequence motifs conserved within phylogenetically related subfamily members.

GPRC5B (also known as Retinoic Acid-Induced Gene 2 Protein (RAIG2)) is an orphan GPCR most closely related to the Class C GPCR family, based on homology. Class C GPCRs also includes the metabotropic glutamate, calcium-sensing receptor, GABAB, and pheromone receptors.

Molecular biology approaches and knockout mouse studies reveal that GPRC5 family proteins have pivotal roles in cancer progression and control of metabolic homeostasis pathways[2].

GPCR5B is known to be involved in the regulation of brain homeostasis, proteins related to adhesion or signaling, and remarkably, GPRC5B-mediated tyrosine-phosphorylation signaling cascades play a critical role in development of obesity and insulin resistance through dynamic sphingolipid metabolism[3].

GPRC5B contains multiple phosphorylated residues in its carboxyl terminus. Phosphorylation of GPRC5b by the tyrosine kinase Fyn and the subsequent direct interaction with the Fyn Src homology 2 (SH2) domain were found to be critical for the initiation and progression of inflammatory signaling in adipose tissue[4]. For example, a GPRC5B mutant lacking the direct binding site for Fyn failed to activate a positive feedback loop of nuclear factor κB-inhibitor of κB kinase ε signaling.

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: 08/01/2023

Anti-GPRC5B Antibody (#AGR-072) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize GPRC5B from rat, mouse and human samples.

For research purposes only, not for human use
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