Periostin ELISA Assay Kit

Additional Information

Assay Background

Periostin (OSF-2) is secreted as a 91 kDa homodimeric soluble extracellular matrix protein expressed in collagen-rich fibrous connective tissues. Periostin is involved in osteoblast recruitment, attachment and spreading. It has been associated with the epithelial-mesenchymal transition in cancer and with the differentiation of mesenchyme in the developing heart. Periostin has functions in osteology, tissue repair, oncology, cardiovascular and respiratory diseases, and in various inflammatory settings. There are at least 7 isoforms of Periostin, caused by alternative splicing (https://www.uniprot.org/uniprot/Q15063).

Assay Procedure

1. Add 150 µl pre-diluted (1+50) STD/CTRL/SAMPLE (see chapter 5) reagents and sample preparation) into each well, swirl gently.
2. Cover tightly and incubate for 2 hours at room temperature (18-26°C).
3. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer). After final wash, remove remaining WASHBUF by strongly tapping plate against paper towel.
4. Add 150 µl AB (biotinylated anti periostin antibody, green cap) into each well, swirl gently.
5. Cover tightly and incubate for 2 hours at room temperature (18-26°C).
6. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer). After final wash, remove remaining WASHBUF by strongly tapping plate against paper towel.
7. Add 150 µl CONJ (Conjugate, amber cap) into each well, swirl gently.
8. Cover tightly and incubate for 1 hour at room temperature (18-26°C).
9. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer). After final wash, remove remaining WASHBUF by strongly tapping plate against paper towel.
10. Add 150 µl SUB (Substrate, blue cap) into each well, swirl gently.
11. Incubate for 30 min at room temperature (18-26°C) in the dark.
12. Add 50 µl STOP (Stop solution, white cap) into each well, swirl gently.
13. Measure absorbance immediately at 450 nm with reference 630 nm, if available.

Expected Values

Values from apparently healthy individuals:

• Median serum (n=24): 864 pmol/l
• Median EDTA plasma (n=20): 817 pmol/l
• Median heparin plasma (n=20): 891 pmol/l
• Median citrate plasma (n=24): 885 pmol/l

It is recommended to establish the normal range for each laboratory.

Literature

• The role of periostin in tissue remodeling across health and disease. Conway SJ et al., Cell Mol Life Sci, 2014; 71(7): 1279-1288.
• Serum periostin is associated with fracture risk in postmenopausal women: a 7-year prospective analysis of the OFELY study. Rousseau JC et al., J Clin Endocrinol Metab, 2014; 2533-2539.
• Circulating periostin levels in patients with AS: association with clinical and radiographic variables, inflammatory markers and molecules involved in bone formation. Sakellariou GT et al., Rheumatology, 2015; 54: 908-914.
• Serum Periostin: A Novel Biomarker for Asthma Management. Hisako Matsumoto, Allergology International, 2014; 63:153-160.
• Serum periostin in obstructive airways disease. Fingleton J et al., Eur Respir J 2016; 44. Doi 10.1183/13993003.01384-2015.
• Levels of Blood Periostin Decrease After Acute Myocardial Infarction and Are Negatively Associated With Ventricular Function After 3 Months. Cheng CW et al., J Investig Med, 2015; 60: 523-528.
• Discoidin domain receptor-1 and periostin: new players in chronic kidney disease. Alfieri C et al, Nephrol Dial Transplant, 2015; 30: 1965-1971.
• Overexpression of periostin in stroma positively associated with aggressive prostate cancer. Tian Y et al., PLoS One, 2015; 17; 10(3):e0121502.
• Role of periostin in cancer progression and metastasis: inhibition of breast cancer progression and metastasis by anti-periostin antibody in a murine model. Int J Mol Med, 2011; 28(2):181-186.
• Tissue expression and serum levels of periostin during pregnancy: A new biomarker of embryo-endometrial cross talk at implantation. Morelli M et al., PLoS One, 2015; 17;10(3):e0121502.