Osteoactivin (also named GPNMB and DC-HIL) is a 125 kDa, intracellular glycoprotein that is associated with cell endosomal/lysosomal compartments (1, 2). Mouse osteoactivin is synthesized as a type I, transmembrane, 574 amino acid (aa) precursor that contains a 22 aa signal sequence, a 478 aa luminal/extracellular domain, a 23 aa transmembrane segment and a 51 aa cytoplasmic tail. The luminal region contains an N-terminal heparin-binding motif, multiple glycosylation sites, an RGD motif and a 130 aa PKD domain. The intracellular tail also has an RGD motif, plus an ITAM (Y-x-x-I) and lysosomal targeting (L-L) motif. The extracellular/luminal region is 89% and 74% aa identical to the equivalent regions in rat and human, respectively. Cells known to express osteoactivin include osteoblasts, dendritic cells, and melanocytes, plus fetal chondrocytes and stratum basale keratinocytes (2, 3). Osteoactivin is reported to bind to heparan sulfate-proteoglycan, possibly on the surface of fibroblasts and endothelial cells (2). It may also interact with integrins.
Key Product Details
Species Reactivity
Mouse
Applications
Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Rat IgG2B Clone # 297310
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Product Specifications
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse Osteoactivin/GPNMB
Lys23-Asn502
Accession # NP_444340
Lys23-Asn502
Accession # NP_444340
Specificity
Detects mouse Osteoactivin/GPNMB in direct ELISAs and Western blots. In direct ELISAs and Western blots, this antibody does not cross-react with recombinant human Osteoactivin.
Clonality
Monoclonal
Host
Rat
Isotype
IgG2B
Scientific Data Images for Mouse Osteoactivin/GPNMB Antibody
Detection of Osteoactivin/GPNMB by Immunohistochemistry
Features of DBA-g+ and DBA mice.(A) Expression of Gpnmb is observed in DBA-g+ but not DBA mice following a single injection of CCl4. However, (B) sequential changes in serum ALT levels and the degree of liver injury are not affected by the lack of Gpnmb-positive macrophages (original magnification, x100). Conversely, (C) the areas of fibrosis and (D) the number of alpha -SMA-positive cells are significantly decreased in the liver tissues of DBA mice compared to DBA-g+ mice (original magnification, x200). Additionally, (E) although lack of Gpnmb-positive macrophages does not affect expression of TGF-beta or Col1 alpha 1, expression of MMP-9, MMP-13, and TIMP-1 are significantly decreased in mice lacking Gpnmb expression at six or eight days after single injection of CCl4. Values are mean ± SEM (n = 4). * P < 0.05 (Mann-Whitney U test). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26599547), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Osteoactivin/GPNMB by Western Blot
Features of DBA-g+ and DBA mice.(A) Expression of Gpnmb is observed in DBA-g+ but not DBA mice following a single injection of CCl4. However, (B) sequential changes in serum ALT levels and the degree of liver injury are not affected by the lack of Gpnmb-positive macrophages (original magnification, x100). Conversely, (C) the areas of fibrosis and (D) the number of alpha -SMA-positive cells are significantly decreased in the liver tissues of DBA mice compared to DBA-g+ mice (original magnification, x200). Additionally, (E) although lack of Gpnmb-positive macrophages does not affect expression of TGF-beta or Col1 alpha 1, expression of MMP-9, MMP-13, and TIMP-1 are significantly decreased in mice lacking Gpnmb expression at six or eight days after single injection of CCl4. Values are mean ± SEM (n = 4). * P < 0.05 (Mann-Whitney U test). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26599547), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Osteoactivin/GPNMB by Immunohistochemistry
Sequential changes in and localization of Gpnmb expression. Gpnmb expression is enhanced in the recovery phase (A) in whole liver as determined by Western blotting and (B) in isolated hepatic macrophages as determined by quantitative real-time polymerase chain reaction (n = 5 at each day). (C) Moreover, Gpnmb expression is observed immunohistochemically around injured lesions in a pattern similar to that in F4/80-positive macrophages, and Gpnmb-positive cells exhibit partial phagocytosis. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26599547), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Osteoactivin/GPNMB by Immunohistochemistry
Characteristics of gpnmb-positive cells infiltrating liver tissue. In LMNCs isolated from injured liver, (A) more than 90% of hepatic macrophages positive for F4/80 are also positive for CD68, whereas CD11b is expressed in approximately 10% of F4/80-positive cells. Gpnmb expression is detected in approximately 50% of CD68-positive cells. (B) Immunofluorescent double staining shows that some CD68-positive cells are also positive for Gpnmb (scale bar: upper, 100 μm; lower, 50 μ m). (C) Some Gpnmb-positive cells show phagocytosis of apoptotic cells (scale bar: left side, 100 μm; right side, 50 μm). (D) Phagocytic activity of CD68-positive cells is significantly higher than that of CD11b-positive cells. Additionally, in CD68-positive cells, phagocytic activity of Gpnmb-positive cells is significantly increased compared with that in Gpnmb-negative cells. Values are mean ± SEM (n = 3). * P < 0.05 (Mann-Whitney U test). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26599547), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Osteoactivin/GPNMB by Western Blot
Sequential changes in and localization of Gpnmb expression. Gpnmb expression is enhanced in the recovery phase (A) in whole liver as determined by Western blotting and (B) in isolated hepatic macrophages as determined by quantitative real-time polymerase chain reaction (n = 5 at each day). (C) Moreover, Gpnmb expression is observed immunohistochemically around injured lesions in a pattern similar to that in F4/80-positive macrophages, and Gpnmb-positive cells exhibit partial phagocytosis. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26599547), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Mouse Osteoactivin/GPNMB Antibody
Application
Recommended Usage
Western Blot
1 µg/mL
Sample: Recombinant Mouse Osteoactivin/GPNMB Fc Chimera (Catalog # 2330-AC)
Sample: Recombinant Mouse Osteoactivin/GPNMB Fc Chimera (Catalog # 2330-AC)
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
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.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.
Calculators
Background: Osteoactivin/GPNMB
References
- Bachner, D. et al. (2002) Gene Exp. Patterns 1:159.
- Shikano, S. et al. (2001) J. Biol. Chem. 276:8125.
- Owen, T.A. et al. (2003) Crit. Rev. Eukaryot. Gene Expr. 13:205.
Long Name
Glycoprotein Non-Metastatic Melanoma Protein B
Alternate Names
DC-HIL, GPNMB, HGFIN
Gene Symbol
GPNMB
UniProt
Additional Osteoactivin/GPNMB Products
Product Documents for Mouse Osteoactivin/GPNMB Antibody
Certificate of Analysis
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Product Specific Notices for Mouse Osteoactivin/GPNMB Antibody
For research use only
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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