Click here to view cell apoptosis assay products

Human OPN(Osteopontin) ELISA Kit

  • Cat.No.:E-EL-H1347

  • Reactivity: Human

To Purchase E-EL-H1347

  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $609

Product Details


Assay type Sandwich-ELISA
Format 96T/48T
Assay time 3.5h
Detection range 0.31-20 ng/mL
Sensitivity 0.19 ng/mL
Sample type &Sample volume serum, plasma and other biological fluids; 100μL
Specificity This kit recognizes Human OPN in samples. No significant cross-reactivity or interference between Human OPN and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of Human OPN concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human OPN. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human OPN and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human OPN, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human OPN. You can calculate the concentration of Human OPN in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips
-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8°C
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data


Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human OPN were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human OPN were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 0.97 2.46 7.02 1.05 2.26 6.75
Standard deviation 0.06 0.14 0.28 0.05 0.11 0.23
CV (%) 6.19 5.69 3.99 4.76 4.87 3.41


The recovery of Human OPN spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 88-100 95
EDTA plasma (n=8) 97-111 102
Cell culture media (n=8) 85-95 90


Samples were spiked with high concentrations of Human OPN and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database LinksSwissProt:   P10451
SynonymsSPP1, BNSP, BSP1, ETA-1, Bone Sialoprotein I, Secreted Phosphoprotein 1, Early T-Lymphocyte Activation 1
Research AreaCancer, Cardiovascular, Signal transduction, Stem cells

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C

elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C

elisa assay procedure 3

3. Aspirate and wash the plate for 3 times

elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 6

6. Add 50μL Stop Solution

elisa assay procedure 7

7. Read the plate at 450nm immediately. Calculation of the results


  1. Frontiers in Endocrinology (2022) IF: 5.555
    Hyaluronic Acid Correlates With Bone Metastasis and Predicts Poor Prognosis in Small-Cell Lung Cancer Patients

    DOI: 10.3389/fendo.2021.785192

    Sample: plasma
  2. Clinical Lung Cancer (2020) IF: 3.55
    Randomized Study of Maintenance Pemetrexed Versus Observation for Treatment of Malignant Pleural Mesothelioma: CALGB 30901

    DOI: 10.1016/j.cllc.2020.06.025

    Sample: Serum
  3. Advances in Medical Sciences (2022) IF: 2.852
    Evaluation of IGFBP5 expression and plasma osteopontin level in COVID-19 patients

    DOI: 10.1016/j.advms.2022.11.001

    Sample: plasma
    Association of serum FGF-23, klotho, fetuin-A, osteopontin, osteoprotegerin and hs-CRP levels with coronary artery disease

    DOI: 10.1080/00365513.2020.1728786

    Sample: serum
  5. Gene Reports (2020)
    Effects of MTHFR genetic polymorphism on inflammatory protein osteopontin in RA patients: A gender based study in North Indian population

    DOI: 10.1016/j.genrep.2020.100737

    Sample: Serum
Show AllShorten


  • Show all
  • Reviews
  • Q&A

S***********rSubmitted [ Feb 11 2024 ]

Asked: How many samples can you analyze with this kit?



adminSubmitted [ Feb 11 2024 ]

Answered: To ensure the accuracy of experimental results, we recommend to do duplicate for each standard and sample, detailed information i'll reply you by mail.


A*******hSubmitted [ Jan 27 2024 ]

Asked: How many samples can be tested in one Osteopontin Elisa Kit



adminSubmitted [ Jan 27 2024 ]

Answered: Hello Arya Shah,we will reply you by email later.

... Show All Show Less

People Also Bought

Apply for Free Shipping 24T ELISA Kit
*Product Name:
*Catalog Number:
*When will you use it?