96-well microplate

Periostin (OSF-2) Human ELISA

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Description

Periostin, also termed osteoblast-specific factor 2 (OSF-2), is a 90-kDa secreted protein that shares a homology with the insect axon guidance protein fasciclin I (1). Periostin is one of four known mammalian genes that contain fasciclin domains with stabilin 1 and 2, as well as TGFβ-Induced Gene-Human clone 3 (BIGH3) (2).

Periostin protein is composed of a signal sequence, four-coiled fasciclin-like repeats, an amino-terminal cysteine-rich region (EMI domain), and heparin-binding domains present in the carboxyl tail. Periostin contains gamma-carboxyglutamate residues that are formed by vitamin K dependent carboxylation (3). These residues are essential for the binding of calcium. Periostin is thought to be involved in osteoblast recruitment, attachment and spreading. It is a component of the extracellular matrix.

The N-terminus part of periostin (up to exon 16) is conserved, while the C-terminal region (comprising exon 17–23) gives rise to different splice isoforms upon alternative splicing. The isoforms have a molecular weight range from 83 kDa to 93 kDa (4). Six different periostin splice isoforms have been reported, but only four of them were sequenced and annotated.

Periostin is expressed during ontogenesis and down regulated in adult except in bones, in collagen-rich fibrous connective tissues subjected to constant mechanical stress, such as periodontal ligament (PDL), heart valves, skin and tendons. Periostin expression is also observed in niches in direct contact with tissue-specific stem cells in mammary gland, bone, and intestine. Periostin has been found to be overexpressed in various type of human tumors including neuroblastoma, head and neck cancer, nasopharyngeal carcinoma, non-small cell lung carcinoma, breast cancer, colon cancer, pancreatic ductal adenocarcinoma and ovarian cancer (5). Isoforms of periostin are over-expressed by stromal cells in several human ovary, breast, colon and brain tumors.

Abnormal expression of periostin is also linked to angiogenesis and metastasis in epithelial tumors. Periostin is expressed by fibroblasts in the normal tissue and in the stroma of the primary tumour. Infiltrating tumour cells need to induce stromal Periostin expression in the secondary target organ to initiate colonization. Periostin is crucial for cancer stem cell maintenance (6). Periostin up-regulation in cancers usually correlates with aggressiveness and/or poor survival.

Type

Sandwich type ELISA

Other names

Periostin, PN, OSF-2, POSTN, Fasciclin-I like, Osteoblast Specific Factor 2, Pleiotrophin, PTN, Heparin-binding growth-associated molecule, HB-GAM, Heparin-binding growth factor 8, HBGF-8, Heparin-binding neurite outgrowth-promoting factor 2, HBNF-2, Heparin-binding brain mitogen, HBBM, NCK.

Principle of Method

This assay is a sandwich Enzyme Linked-Immunosorbent Assay (ELISA) for quantitative determination of human periostin in cell culture supernatants, serum and plasma. A monoclonal antibody specific for periostin has been precoated onto the 96-well microtiter plate. Standards (STD) and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, periostin is recognized by the addition of a biotinylated monoclonal antibody specific for periostin (DET). After removal of excess biotinylated antibody, streptavidine-peroxidase (STREP-HRP) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3’,5,5’-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450 nm after acidification and is directly proportional to the concentration of periostin in the samples.

Sample Types

Serum, Plasma and Cell Culture Supernatant

Sample Volume

Serum and Plasma : ~2 µL (actual amount of sample loaded in well after dilution). 

Standard Curve

Periostin Human ELISA Standard Curve

Calibration Range:  78 pg/ml – 5000 pg/ml

Limit of Detection

15 pg/ml

Intra-assay (Within-Run) C.V.

7.12 %

Inter-assay (Run-to-Run) C.V.

7.62 %

Resources

References to Summary

  • Periostin: novel diagnostic and therapeutic target for cancer: Y. Kudo, et al.; Histopathol. 22, 1167 (2007)
  • The many facets of the matricellular protein periostin during cardiac development, remodeling, and pathophysiology: A. Russel, et al.; Cell Commun. Signal. 3, 275 (2009)
  • Periostin, a member of a novel family of vitamin K-dependent proteins, is expressed by mesenchymal stromal cells: D.L. Coutu, et al.; Biol. Chem. 283, 17991 (2008)
  • Characterization of periostin isoform pattern in non-small cell lung cancer: L. Morra, et al.; Lung Cancer 76, 183 (2012)
  • Role of periostin in cancer progression and metastasis: inhibition of breast cancer progression and metastasis by anti-periostin antibody in a murine model: M. Kyutoku, et al.; J. Mol. Med. 28, 181 (2011)
  • Interactions between cancer stem cells and their niche govern metastatic colonization: I. Malanchi, et al.; Nature 481, 85 (2011)