Detection of Human GPR3 by Western Blot.
Western blot shows lysates of human brain (Alzheimer's disease outer cortex) tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human GPR3 Monoclonal Antibody (Catalog # MAB7429) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for GPR3 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of GPR3 in HEK293 Human Cell Line Transfected with human GRP3 and eGFP by Flow Cytometry.
HEK293 human embryonic kidney cell line transfected with (A) human GRP3 or (B) irrelevant transfectants, and eGFP was stained with Mouse Anti-Human GPR3 Monoclonal Antibody (Catalog # MAB7429) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). Quadrant markers were set based on Mouse IgG2B Flow Cytometry Isotype Control (Catalog # MAB0041, data not shown). View our protocol for Staining Membrane-associated Proteins.
GPR3 in Human Brain.
GPR3 was detected in immersion fixed paraffin-embedded sections of Alzheimer's human brain using Mouse Anti-Human GPR3 Monoclonal Antibody (Catalog # MAB7429) at 15 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-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal cell bodies and processes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
GPR3, previously called ACCA (adenylate cyclase constitutive activator), is 7-transmembrane receptor with potential sites for glycosylation at the N-terminus and serine phosphorylation at the C-terminus. It has been shown to promote cAMP accumulation, which regulates maintenance of meiotic arrest in oocytes. It is also expressed in the brain, where it has been implicated in cAMP-dependent emotional responses and modulation of amyloid-beta peptide generation. The 330 amino acid (aa) human GPR3 contains 86 extracellular aa that share ~83% aa identity with mouse and rat GPR3.
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