GPR158 Antibody (1027604) [Unconjugated]

Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with human GPR158. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human GPR158 Monoclonal Antibody (Catalog # MAB10286) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for GPR158 at approximately 175 kDa (as indicated). GAPDH (MAB5718) is shown as a loading control.This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

HEK293 human embryonic kidney cell line transfected with (A) human GPR158 or (B) irrelevant protein, and eGFP was stained with Mouse Anti-Human GPR158 Monoclonal Antibody (Catalog # MAB10286) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B). Quadrant markers were set based on control antibody staining (MAB002). Staining was performed using our Staining Membrane-associated Proteins protocol.

GPR158 was detected in immersion fixed paraffin-embedded sections of human brain (cortex) tissue using Mouse Anti-Human GPR158 Monoclonal Antibody (Catalog # MAB10286) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western lane view shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with GPR-158 and eGFP, loaded at 0.2 mg/mL. A specific band was detected for GPR158 at approximately 191 kDa (as indicated) using 25 µg/mL of Mouse Anti-Human GPR158 Monoclonal Antibody (Catalog # MAB10286) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, IHC, CyTOF-ready
• Clone: 1027604
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

GPR158 Antibody (1027604) [Unconjugated] Summary

• Immunogen: Human embryonic kidney cell HEK293-derived human GPR158 protein
  Ala24-Gln411
  Accession # Q5T848
• Specificity: Detects human GPR158 in direct ELISAs.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 5-25 ug/mL
  • Simple Western 25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for GPR158 Antibody (1027604) [Unconjugated]

• G protein-coupled receptor 158
• GPR158
• KIAA1136
• KIAA1136FLJ37801
• probable G-protein coupled receptor 158
• RP11-59G22.1

Background

G-protein coupled receptor 158 (GPR158) is a receptor belonging to the Class C GPCR family. It lacks the extracellular Venus flytrap module characteristic of the known members of that family and instead contains two other elements that are not typical of the class: a calcium-binding EGF-like domain and a leucine repeat region (1, 2). The mature extracellular domain of human GPR158 contains 393 amino acids (aa) and shares 89% identity with both mouse and rat GPR158. GPR158 is expressed at the highest level in the brain, but also in a variety of other tissues including retina, spleen, liver and lung (3). GPR158 was originally identified in functional screens linked with biological stress and has been implicated in the osteocalcin effect on cognitive processes in the brain (4, 5), and glaucoma and cancer in the periphery (4, 6).

1. Jingami, H. et al. (2003) Curr. Opin. Neurobiol. 13:271.
2. Bjarnadóttir, T.K et al. (2005) Gene. 362:70.
3. Orlandi, C. et al. (2012) J. Cell Biol. 197:711.
4. Itakura, T. et al. (2019) J. Ocul. Pharmacol. Ther. 35:203.
5. Khrimian, L. et al. (2017) J. Exp. Med. 214:2859.
6. Fenner, A. (2015) Nat. Rev. Urol. 12:182.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • Q5T848()