Human Osteoactivin/GPNMB Antibody

(4 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Osteoactivin/GPNMB in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant mouse Osteoactivin is observed.
  • Source
    Monoclonal Mouse IgG2B Clone # 303822
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Osteoactivin/GPNMB isoform 2
    Ala22-Asn486
    Accession # NP_002501
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    Recombinant Human Osteoactivin/GPNMB Fc Chimera (Catalog # 2550-AC) under non‑reducing conditions only
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    Immersion fixed MG-63 human osteosarcoma cell line
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Intracellular Staining by Flow Cytometry
Detection of Osteoactivin/GPNMB in MG‑63 Human Cell Line by Flow Cytometry. MG‑63 human osteosarcoma cell line was stained with Mouse Anti-Human Osteoactivin/GPNMB Monoclonal Antibody (Catalog # MAB25501, filled histogram) or isotype control antibody (Catalog # MAB004, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B).To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    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.
Background: Osteoactivin/GPNMB

Osteoactivin (also GPNMB and DC-HIL) is a variably glycosylated 75-125 kDa member of the NMB/pMEL-17 family of molecules. It is found in multiple subcellular sites, but is most often associated with the endosomal/lysosomal compartment (1-3). Human Osteoactivin is a 560 amino acid (aa) type I transmembrane protein. Its precursor contains a 21 aa signal sequence, a 465 aa luminal/extracellular domain, a 21 aa transmembrane segment and a 53 aa cytoplasmic tail (4, 5). The luminal region contains an N-terminal heparin-binding motif (aa 23-26), multiple glycosylation sites, an RGD motif (aa 64-66) and an 88 aa PKD domain (aa 240-327). The intracellular tail has an ITAM (Y-x-x-I) and lysosomal targeting (L-L) motif (4, 5). The extracellular/luminal region shares 74% and 77% aa identity with the equivalent regions in mouse and canine, respectively. Multiple isoforms would appear to exist. There is one alternate splice form known that shows a 12 aa insert between aa 339-340 (6). An addtional 206 aa isoform shows a mutation at position # 181 that results in a 26 aa substitution for the C-terminal 380 amino acids (7, 8). This has the potential to be soluble and may represent a counterpart to a secreted isoform of rat Osteoactivin (9). Cells known to express Osteoactivin include macrophages/Kupffer cells, fibroblasts, osteoblasts, myeloid dendritic cells, retinal pigment epithelial cells and melanocytes, plus fetal chondrocytes and stratum basale keratinocytes (3-5, 10-12). In mice, Osteoactivin is reported to bind to heparan sulfate-proteoglycan, possibly on the surface of endothelial cells and may also interact with integrins (13). It also appears to act as an inflammatory suppressor gene, as its expression downregulates the macrophage inflammatory response by inhibiting IL-6 and IL-12 p40 production (3).

  • References:
    1. Bachner, D. et al. (2002) Gene Exp. Patterns 1:159.
    2. Safadi, F.F. et al. (2002) J. Cell. Biochem. 84:12.
    3. Ripoll, V.M. et al. (2007) J. Immunol. 178:6557. 
    4. Owen, T.A. et al. (2003) Crit. Rev. Eukaryot. Gene Expr. 13:205. 
    5. Weterman, M.A.J. et al. (1995) Int. J. Cancer 60:73. 
    6. Kuan, C-T. et al. (2006) Clin. Cancer Res. 12:1970. 
    7. Lennerz, V. et al. (2005) Proc. Natl. Acad. Sci. USA 102:16013.
    8. Genbank Accession # AAH11595.
    9. Abdelmagid, S.M. et al. (2007) J. Cell. Physiol. 210:26.
    10. Haralanova-Ilieva, B. et al. (2005) J. Hepatol. 42:565.
    11. Ahn, J.H. et al. (2002) Blood 100:1742.
    12. Anderson, M.G. et al. (2002) Nat. Genet. 30:81.
    13. Shikano, S. et al. (2001) J. Biol. Chem. 276:8125.
  • Long Name:
    Glycoprotein Non-Metastatic Melanoma Protein B
  • Entrez Gene IDs:
    10457 (Human); 93695 (Mouse)
  • Alternate Names:
    DC-HIL; glycoprotein (transmembrane) nmb; glycoprotein NMB; GPNMB; HGFIN; HGFINglycoprotein nmb-like protein; NMBosteoactivin; Osteoactivin; Transmembrane glycoprotein HGFIN; transmembrane glycoprotein NMB
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Elevation of glycoprotein nonmetastatic melanoma protein B in type 1 Gaucher disease patients and mouse models
    FEBS Open Bio, 2016;6(9):902-13.
    Species: Mouse
    Sample Type: Plasma
    Application: WB
  2. The transcription factor MITF is a critical regulator of GPNMB expression in dendritic cells.
    Authors: Gutknecht M, Geiger J, Joas S, Dorfel D, Salih H, Muller M, Grunebach F, Rittig S
    Cell Commun Signal, 2015;13(0):19.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Glycoprotein non-metastatic melanoma protein B as a predictive prognostic factor in clear-cell renal cell carcinoma following radical nephrectomy.
    Authors: Qin C, Liu Z, Yuan Y, Zhang X, Li H, Zhang C, Xu T, Wang X
    Mol Med Rep, 2014;9(3):851-6.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC - Not specified
  4. GPNMB/OA protein increases the invasiveness of human metastatic prostate cancer cell lines DU145 and PC3 through MMP-2 and MMP-9 activity.
    Authors: Fiorentini C, Bodei S, Bedussi F, Fragni M, Bonini S, Simeone C, Zani D, Berruti A, Missale C, Memo M, Spano P, Sigala S
    Exp Cell Res, 2014;323(1):100-11.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
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