Human Osteoactivin/GPNMB Alexa Fluor® 488-conjugated Antibody

    
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  • 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
    Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
  • Label
    Alexa Fluor 488
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Intracellular Staining by Flow Cytometry
    0.25-1 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 Example
Intracellular Staining by Flow Cytometry
Detection of Osteoactivin/GPNMB in MG-63 HumanOsteosarcoma Cell Line by Flow Cytometry. MG-63 human osteosarcoma cell line wasstained Mouse Anti-Human Osteoactivin/GPNMB Alexa Fluor® 488-conjugatedMonoclonal Antibody (Catalog # FAB25501G) or isotype control antibody(Catalog # IC0041G, open histogram).  To facilitate intracellular staining, cellswere 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
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Protect from light. Do not freeze.
    • 12 months from date of receipt, 2 to 8 °C as supplied.
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
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Isotype Controls
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Mouse IgG2B Alexa Fluor® 488-conjugated Isotype Control

Ctrl IC0041G 3  
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Staining Reagents
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Flow Cytometry Staining Buffer (1X)

Flow FC001 10
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Flow Cytometry Mouse Lyse Buffer (10X)

Flow FC003 5
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Flow Cytometry Human Lyse Buffer (10X)

Flow FC002 1
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