Periostin ELISA, human
- Product Details
- Instructions For Use
- Validation Data
- Additional Information
- References, Applications
|Method:||Sandwich ELISA, HRPO/TMB, 12x8-well strips|
|Sample type:||Serum, plasma (EDTA, heparin, citrate)|
|Standard range:||0-4000 pmol/l (7 serum based standards)|
|Standard points:||0/125/250/500/1000/2000/4000 pmol/l|
|Control:||2 serum based controls|
|Sample size:||10 µl / sample|
|Incubation time:||2 h / 2 h / 1 h / 30 min|
|Unit conversion:||1 pg/ml = 0.011 pmol/l (MW: 91 kDa)|
LOD: 20 pmol/l (0 pmol/l + 3 SD); LLOQ: 62.5 pmol/l
Intra-assay: 2 samples of known concentrations were tested 5 times within 1 kit lot by 1 operator.
Inter-assay: 2 samples of known concentrations were tested 10 times within 3 different kit lots by 3 different operators.
|Intra-assay (n=5)||Sample 1||Sample 2|
|Inter-assay (n=10)||Sample 1||Sample 2|
The recovery of Periostin was evaluated by adding 2 known concentrations of human recombinant Periostin to different human sample matrices.
|Matrix||Mean S/R [% ]|
|+500 pmol/l||+2000 pmol/l|
|EDTA plasma (n=8)||98||83|
|Heparin plasma (n=7)||92||85|
|Citrate plasma (n=8)||102||91|
Dilution linearity was assessed by serially diluting samples containing endogenous Periostin with assay buffer.
|Matrix||Mean R of dilution [% ]|
|EDTA plasma (n=4)||99||115|
|Heparin plasma (n=4)||96||126|
|Citrate plasma (n=4)||95||122|
This assay is optimized to detect all known splicing forms of human periostin. This assay recognizes recombinant and endogenous Periostin.
Due to the high sequence homology between human Periostin and Periostin of other species, the antibodies utilized in the assay may cross-react with mouse, rat, cynomolgous monkey, dog and cat Periostin.
This immunoassay is calibrated against recombinant human Periostin peptide.
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.
Principle of the assay:
Manual ELISAs - easily adaptable for automation!
INSTRUCTIONS FOR USE
Click link for:
Click here for assay validation data (S/R, dilution linearity, precision, ...).
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 (http://www.uniprot.org/uniprot/Q15063).
Click here for the Periostin ELISA reference list.
Characterization of a sandwich ELISA for the quantification of all human periostin isoforms.
Gadermaier E, Tesarz M, Suciu AA, Wallwitz J, Berg G, Himmler G.J
Clin Lab Anal. 2018 Feb;32(2).
Effect of age and gender on serum periostin: Relationship to cortical measures, bone turnover and hormones.
Walsh JS, Gossiel F, Scott JR, Paggiosi MA, Eastell R.
Bone. 2017 Jun;99:8-13.
ASBMR Annual Meeting 2016, Sept 16-19
Georgia World Congress Center, Atlanta, GA, USA
Abstract of poster # LB-SU0353
Novel ELISA for the measurement of human Periostin
Manfred Tesarz, Elisabeth Gadermaier, Gabriela Berg, Gottfried Himmler
The Antibody Lab GmbH, Vienna, Austria
Purpose: Periostin (osteoblast-specific factor OSF-2) is a component of the extracellular matrix and is thought to be involved in osteoblast recruitment, attachment and spreading. As a potential biomarker of bone turnover it may assist in the management of bone diseases. Periostin consists of a conserved N-terminus and a C-terminal region which is affected by alternative splicing. Currently, at least seven splicing isoforms of human Periostin have been identified.
Methods: We developed a sandwich ELISA, which enables the detection of all known human circulating Periostin isoforms. Our novel assay utilizes monoclonal and affinity-purified polyclonal antibodies and recognizes epitopes that are conserved between human and animal species, e.g. mouse, rat, cynomolgus macaque, dog, and cat Periostin.
Results: The novel Periostin ELISA assay is optimized for human serum and plasma and covers a wide calibration range between 125 to 4,000 pmol/l. Assay characteristics, such as precision (intra-assay: ≤3%, inter-assay: ≤6%), dilution linearity (99-115%) and spike-recovery (83 – 106%), the matrix comparison between serum and EDTA-plasma (R2 0,96) as well as sample stability meet the standards of acceptance. Periostin serum and plasma concentrations in apparently healthy individuals are 864 +/- 269 pmol/l (n=24) and 817 +/- 170 pmol/l (n=20), respectively.
Conclusion: This ELISA provides a reliable and accurate tool for the quantitative determination of Periostin in human healthy and diseased samples.