Human SPARC Antibody

(7 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human SPARC/Osteonectin in direct ELISAs and Western blots. In direct ELISAs, approximately 25% cross-reactivity with recombinant mouse SPARC is observed, and less than 3% cross-reactivity with recombinant human SPARC L1 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human SPARC/Osteonectin
    Ala18-Ile303
    Accession # P09486
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Simple Western
    20 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    MG‑63 human osteosarcoma cell line fixed with paraformaldehyde and permeabilized with saponin
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 SPARC by Western Blot. Western blot shows lysates of MG‑63 human osteosarcoma cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human SPARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF941) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for SPARC at approximately 43 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

SPARC/Osteonectin in Human Ovary. SPARC/Osteonectin was detected in immersion fixed paraffin-embedded sections of human ovary using Goat Anti-Human SPARC/Osteonectin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF941) 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). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human SPARC by Simple WesternTM. Simple Western lane view shows lysates of MG‑63 human osteosarcoma cell line, loaded at 0.2 mg/mL. A specific band was detected for SPARC at approximately 57 kDa (as indicated) using 20 µg/mL of Goat Anti-Human SPARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF941) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.

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: SPARC

SPARC, an acronym for “secreted protein, acidic and rich in cysteine”, is also known as osteonectin or BM-40 (1-5). It is the founding member of a family of secreted matricellular proteins with similar domain structure. The 286 amino acid (aa), 43 kDa protein contains an N-terminal acidic region that binds calcium, a follistatin domain that contains Kazal-like sequences, and a C-terminal extracellular calcium (EC) binding domain with two EF-hand motifs (1-5). Crystal structure modeling shows that residues implicated in cell binding, inhibition of cell spreading, and disassembly of focal adhesions cluster on one face of SPARC, while a collagen binding epitope and an N-glycosylation site are opposite this face (6). SPARC is produced by fibroblasts, capillary endothelial cells, platelets and macrophages, especially in areas of tissue morphogenesis and remodeling (3, 7). SPARC shows context-specific effects, but generally inhibits adhesion, spreading and proliferation, and promotes collagen matrix formation (3-5). For endothelial cells, SPARC disrupts focal adhesions and binds and sequesters PDGF and VEGF (3-5). SPARC is abundantly expressed in bone, where it promotes osteoblast differentiation and inhibits adipogenesis (5, 8). SPARC is potentially cleaved by metalloproteinases, producing an angiogenic peptide that includes the copper-binding sequence KGHK (7). Paradoxically, SPARC is highly expressed in many tumor types undergoing an endothelial to mesenchymal transistion; its expression, however, mainly decreases the likelihood of metastasis and confers sensitivity to chemotherapy and radiation (4, 9-11). Stabilin-1, which is expressed on alternately activated macrophages, is the first SPARC receptor to be identified. It binds the SPARC EC domain and mediates endocytosis for degradation (12). Mature human SPARC shows 92%, 92%, 97%, 99%, 96%, and 85% aa identity with mouse, rat, canine, bovine, porcine, and chick SPARC, respectively.

  • References:
    1. Lankat-Buttgereit, B. et al. (1988) FEBS Lett. 236:352.
    2. Sweetwyne, M.T. et al. (2004) J. Histochem. Cytochem. 52:723.
    3. Sage, H. et al. (1989) J. Cell Biol. 109:341.
    4. Framson, P.E. and E.H. Sage (2004) J. Cell. Biochem. 92:679.
    5. Alford, A.I. and K. D. Hankenson (2006) Bone 38:749.
    6. Hohenester, E et al. (1997) EMBO J. 16:3778.
    7. Sage, E.H. et al. (2003) J. Biol. Chem. 278:37849.
    8. Delany, A.M. et al. (2003) Endocrinology 144:2588.
    9. Robert, G. et al. (2006) Cancer Res. 66:7516.
    10. Koblinski, J.E. et al. (2005) Cancer Res. 65:7370.
    11. Tai, I.T. et al. (2005) J. Clin. Invest. 115:1492.
    12. Kzhyshkowska, J. et al. (2006) J. Immunol. 176:5825.
  • Long Name:
    Secreted Protein Acidic and Rich in Cysteine
  • Entrez Gene IDs:
    6678 (Human); 20692 (Mouse)
  • Alternate Names:
    Basement-membrane protein 40; BM-40; ONcysteine-rich protein; Osteonectin; Secreted protein acidic and rich in cysteine; secreted protein, acidic, cysteine-rich (osteonectin); SPARC
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.

7 Citations: Showing 1 - 7
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Species
Applications
Sample Type
  1. Stabilin-1 is expressed in human breast cancer and supports tumor growth in mammary adenocarcinoma mouse model
    Oncotarget, 2016;7(21):31097-110.
  2. Automated Analysis and Classification of Histological Tissue Features by Multi-Dimensional Microscopic Molecular Profiling.
    Authors: Riordan D, Varma S, West R, Brown P
    PLoS ONE, 2015;10(7):e0128975.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  3. Collagen signaling enhances tumor progression after anti-VEGF therapy in a murine model of pancreatic ductal adenocarcinoma.
    Authors: Aguilera K, Rivera L, Hur H, Carbon J, Toombs J, Goldstein C, Dellinger M, Castrillon D, Brekken R
    Cancer Res, 2014;74(4):1032-44.
    Species: Human
    Sample Type: Recombinant Protein
    Application: ELISA Development
  4. Targeting SPARC by lentivirus-mediated RNA interference inhibits cervical cancer cell growth and metastasis.
    BMC Cancer, 2012;12(0):464.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  5. SPARC is a key regulator of proliferation, apoptosis and invasion in human ovarian cancer.
    Authors: Chen, Jie, Wang, Mei, Xi, Bo, Xue, Jian, He, Dan, Zhang, Jie, Zhao, Yueran
    PLoS ONE, 2012;7(8):e42413.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  6. Diverse functions of reactive cysteines facilitate unique biosynthetic processes of aggregate-prone interleukin-31.
    Authors: Shen M, Siu S, Byrd S, Edelmann KH, Patel N, Ketchem RR, Mehlin C, Arnett HA, Hasegawa H
    Exp. Cell Res., 2011;317(7):976-93.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  7. SPARC is a VCAM-1 counter-ligand that mediates leukocyte transmigration.
    Authors: Kelly KA, Allport JR, Yu AM, Sinh S, Sage EH, Gerszten RE, Weissleder R
    J. Leukoc. Biol., 2007;81(3):748-56.
    Species: Human
    Sample Type: Protein
    Application: WB
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