Periostin ELISA Kit

Catalog No. ABIN422957

Product Details Periostin ELISA Kit

Target: Periostin (POSTN)

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 78 pg/mL - 5000 pg/mL

Minimum Detection Limit: 78 pg/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of POSTN in Serum,Plasma,Biological Fluids

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Specificity:

This assay has high sensitivity and excellent specificity for detection of Periostin (POSTN).
No significant cross-reactivity or interference between Periostin (POSTN) and analogues was observed.

Cross-Reactivity (Details): No significant cross-reactivity or interference between Periostin (POSTN) and analogues was observed.

Sensitivity: 31 pg/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Material not included:

• Microplate reader with 450 nm filter.
• Precision single or multi-channel pipettes and disposable tips.
• Eppendorf Tubes for diluting samples.
• Deionized or distilled water.
• Absorbent paper for blotting the microtiter plate.
• Container for Wash Solution

Application Details

Application Notes:

• Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
• The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
• Kits from different batches may be a little different in detection range, sensitivity and color developing time.
• Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
• Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
• There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
• Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
• Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
• Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
• Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Periostin (POSTN). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Periostin (POSTN). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Periostin (POSTN), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Periostin (POSTN) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 0.5 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 5,000pg/mL. Prepare 7 tubes containing 0.25 mL Standard Diluent and produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 5,000pg/mL, 2,500pg/mL, 1,250pg/mL, 625pg/mL, 312pg/mL, 156pg/mL, 78pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Sample Collection: Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.

Calculation of Results:

Average the duplicate readings for each standard, control, and samples and subtract the average zero standard optical density. Construct a standard curve by plotting the mean O.D. and concentration for each standard and draw a best fit curve through the points on the graph or create a standard curve on log-log graph paper with POSTN concentration on the y-axis and absorbance on the x-axis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the known concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. However, the O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), plotting log of the data to establish standard curve for each test is recommended. Typical standard curve below is provided for reference only.

Assay Precision:

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Periostin (POSTN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Periostin (POSTN) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Handling Advice: The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Storage: 4 °C

Storage Comment:

• For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
• For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
  Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
• For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.

Expiry Date: 6 months

References for Periostin ELISA Kit (ABIN422957)

Chen, Huo, Ren, Li, Sun, Li, Zhang, Chen, Song: "Polycystic Ovary Syndrome is Associated with Elevated Periostin Levels." in: Experimental and clinical endocrinology & diabetes : official journal, German Society of Endocrinology [and] German Diabetes Association, Vol. 127, Issue 9, pp. 571-577, (2020) (PubMed).

Guañabens, Filella, Florez, Ruiz-Gaspá, Conesa, Peris, Monegal, Torres: "Tartrate-resistant acid phosphatase 5b, but not periostin, is useful for assessing Paget's disease of bone." in: Bone, Vol. 124, pp. 132-136, (2020) (PubMed).

Wang, Fu, Shen, Zhang, Wang, Wu: "Identification and diagnostic value of pleural fluid periostin and serum periostin of malignant pleural effusions in patients with non-small-cell lung cancer." in: Journal of clinical laboratory analysis, Vol. 33, Issue 7, pp. e22943, (2020) (PubMed).

Rousseau, Bertholon, Chapurlat, Szulc: "Serum periostin is associated with cancer mortality but not cancer risk in older home-dwelling men: A 8-year prospective analysis of the STRAMBO study." in: Bone, Vol. 132, pp. 115184, (2020) (PubMed).

Pepe, Bonnet, Herrmann, Biver, Rizzoli, Chevalley, Ferrari: "Interaction between LRP5 and periostin gene polymorphisms on serum periostin levels and cortical bone microstructure." in: Osteoporosis international : a journal established as result of cooperation between the European Foundation for Osteoporosis and the National Osteoporosis Foundation of the USA, Vol. 29, Issue 2, pp. 339-346, (2019) (PubMed).

Polyzos, Kountouras, Anastasilakis, Papatheodorou, Kokkoris, Terpos: "Circulating periostin in patients with nonalcoholic fatty liver disease." in: Endocrine, Vol. 56, Issue 2, pp. 438-441, (2018) (PubMed).

Walsh, Gossiel, Scott, Paggiosi, Eastell: "Effect of age and gender on serum periostin: Relationship to cortical measures, bone turnover and hormones." in: Bone, Vol. 99, pp. 8-13, (2018) (PubMed).

Gadermaier, Tesarz, Suciu, Wallwitz, Berg, Himmler: "Characterization of a sandwich ELISA for the quantification of all human periostin isoforms." in: Journal of clinical laboratory analysis, Vol. 32, Issue 2, (2018) (PubMed).

Dong, Yu, Du, Wang, Zhu, Yang, Che, Shen, Jiang: "Serum periostin concentrations and outcomes after severe traumatic brain injury." in: Clinica chimica acta; international journal of clinical chemistry, Vol. 471, pp. 298-303, (2018) (PubMed).

Garnero, Bonnet, Ferrari: "Development of a New Immunoassay for Human Cathepsin K-Generated Periostin Fragments as a Serum Biomarker for Cortical Bone." in: Calcified tissue international, Vol. 101, Issue 5, pp. 501-509, (2018) (PubMed).

Bonnet, Biver, Chevalley, Rizzoli, Garnero, Ferrari: "Serum Levels of a Cathepsin-K Generated Periostin Fragment Predict Incident Low-Trauma Fractures in Postmenopausal Women Independently of BMD and FRAX." in: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, Vol. 32, Issue 11, pp. 2232-2238, (2018) (PubMed).

Ji, Chou, Wu, Shen, Yang, Wang, Lan, Shi: "Association between serum periostin concentrations and outcome after acute spontaneous intracerebral hemorrhage." in: Clinica chimica acta; international journal of clinical chemistry, Vol. 474, pp. 23-27, (2018) (PubMed).

Ding, Ge, Qu, Feng, Long, Wei, Zhou, Wu, Yao, Deng: "Increased serum periostin concentrations are associated with the presence of diabetic retinopathy in patients with type 2 diabetes mellitus." in: Journal of endocrinological investigation, Vol. 41, Issue 8, pp. 937-945, (2018) (PubMed).

Yan, Liu, Li, Chen, Bian, Zhao, Han, Han, Han, Wang, Yang: "Circulating periostin levels increase in association with bone density loss and healing progression during the early phase of hip fracture in Chinese older women." in: Osteoporosis international : a journal established as result of cooperation between the European Foundation for Osteoporosis and the National Osteoporosis Foundation of the USA, Vol. 28, Issue 8, pp. 2335-2341, (2017) (PubMed).

Anastasilakis, Polyzos, Tsoli, Papatheodorou, Kokkoris, Kaltsas, Terpos, Makras: "Low periostin levels in adult patients with Langerhans cell histiocytosis are independently associated with the disease activity." in: Metabolism: clinical and experimental, Vol. 71, pp. 198-201, (2017) (PubMed).

Garnero: "The Utility of Biomarkers in Osteoporosis Management." in: Molecular diagnosis & therapy, Vol. 21, Issue 4, pp. 401-418, (2017) (PubMed).

Zhang, Yuan, Yao, Sun, Shi, Su: "Predictive and prognostic value of serum periostin in advanced non-small cell lung cancer patients receiving chemotherapy." in: Tumour biology, Vol. 39, Issue 5, pp. 1010428317698367, (2017) (PubMed).

Zhu, Zhu, Dai, Li, Fang, Zhao, Wan, Wang, Wang, Yu, Li: "Serum periostin is a potential biomarker for non-alcoholic fatty liver disease: a case-control study." in: Endocrine, Vol. 51, Issue 1, pp. 91-100, (2016) (PubMed).

Terpos, Christoulas, Kastritis, Bagratuni, Gavriatopoulou, Roussou, Papatheodorou, Eleutherakis-Papaiakovou, Kanellias, Liakou, Panagiotidis, Migkou, Kokkoris, Moulopoulos, Dimopoulos: "High levels of periostin correlate with increased fracture rate, diffuse MRI pattern, abnormal bone remodeling and advanced disease stage in patients with newly diagnosed symptomatic multiple myeloma." in: Blood cancer journal, Vol. 6, Issue 10, pp. e482, (2016) (PubMed).

Rousseau, Sornay-Rendu, Bertholon, Garnero, Chapurlat: "Serum periostin is associated with prevalent knee osteoarthritis and disease incidence/progression in women: the OFELY study." in: Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society, Vol. 23, Issue 10, pp. 1736-42, (2015) (PubMed).

Target Details for Periostin

Target: Periostin (POSTN)

Alternative Name: POSTN (POSTN Products)

Synonyms: fa99h07 ELISA Kit, pn ELISA Kit, postn ELISA Kit, wu:fa99h07 ELISA Kit, wu:fc70f09 ELISA Kit, zgc:153873 ELISA Kit, LOC100008945 ELISA Kit, osf-2 ELISA Kit, pdlpostn ELISA Kit, periostin ELISA Kit, OSF-2 ELISA Kit, OSF2 ELISA Kit, PDLPOSTN ELISA Kit, PN ELISA Kit, RP11-412K4.1 ELISA Kit, A630052E07Rik ELISA Kit, AI747096 ELISA Kit, Osf2 ELISA Kit, PLF ELISA Kit, peri ELISA Kit, Plf ELISA Kit, periostin ELISA Kit, periostin, osteoblast specific factor b ELISA Kit, periostin, osteoblast specific factor ELISA Kit, POSTN ELISA Kit, postnb ELISA Kit, postn ELISA Kit, Postn ELISA Kit

UniProt: Q15063