Human Osteoactivin/GPNMB Antibody

(6 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Osteoactivin in ELISAs and Western blots. In sandwich immunoassays, less than 2% cross-reactivity with recombinant mouse Osteoactivin is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Osteoactivin isoform b
    Lys23-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
    0.5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
    • Human Osteoactivin/GPNMB Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Human Osteoactivin/GPNMB Antibody (Catalog # AF2550)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human Osteoactivin/GPNMB Biotinylated Antibody (Catalog # BAF2550)
  • ELISA Standard
     
    Recombinant Human Osteoactivin/GPNMB Fc Chimera Protein, CF (Catalog # 2550-AC)
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
Detection of Human Osteoactivin/GPNMB by Western Blot. Western blot shows lysates of U‑118‑MG human glioblastoma/astrocytoma cell line and T98G human glioblastoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Osteoactivin/GPNMB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2550) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for Osteoactivin/GPNMB at approximately 95 and 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
Osteoactivin/GPNMB in Human Liver. Osteoactivin/GPNMB was detected in immersion fixed paraffin-embedded sections of human liver using Goat Anti-Human Osteoactivin/GPNMB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2550) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to Kupffer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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.

6 Citations: Showing 1 - 6
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Species
Applications
Sample Type
  1. Exploratory study on the effect of osteoactivin on muscle regeneration in a rat volumetric muscle loss model
    Authors: J Ma, AR Baker, A Calabro, KA Derwin
    PLoS ONE, 2017;12(4):e0175853.
    Species: N/A
    Sample Type: Gel Constructs
    Application: IHC Frozen
  2. Identification of a biomarker in cerebrospinal fluid for neuronopathic forms of Gaucher disease.
    Authors: Zigdon, Hila, Savidor, Alon, Levin, Yishai, Meshcheriakova, Anna, Schiffmann, Raphael, Futerman, Anthony
    PLoS ONE, 2015;10(3):e0120194.
    Species: Human
    Sample Type: CSF
    Application: WB
  3. 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: Cell Lysates
    Application: WB
  4. Hematopoietic growth factor inducible neurokinin-1 (Gpnmb/Osteoactivin) is a biomarker of progressive renal injury across species.
    Authors: Patel-Chamberlin M, Wang Y, Satirapoj B
    Kidney Int., 2011;79(10):1138-48.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  5. Inactivation of the FLCN tumor suppressor gene induces TFE3 transcriptional activity by increasing its nuclear localization.
    Authors: Hong SB, Oh H, Valera VA, Baba M, Schmidt LS, Linehan WM
    PLoS ONE, 2010;5(12):e15793.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Glycoprotein nonmetastatic B is an independent prognostic indicator of recurrence and a novel therapeutic target in breast cancer.
    Authors: Rose AA, Grosset AA, Dong Z, Russo C, Macdonald PA, Bertos NR, St-Pierre Y, Simantov R, Hallett M, Park M, Gaboury L, Siegel PM
    Clin. Cancer Res., 2010;16(7):2147-56.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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