Osteopontin (OPN), also known as bone sialoprotein (BSP), is a secreted SIBLING family protein that can be variably modified by O- and N-glycosylation, sulfation, phosphorylation, and transglutamination. OPN is widely expressed and is prominent in mineralized tissues. It inhibits bone mineralization and kidney stone formation and promotes inflammation, cell adÂhesion, and migration. Its expression is upregulatÂed during inflammation, obesity, atherosclerosis, cancer, and tissue damage and contributes to the pathophysiology of these conditions. The central region of OPN contains RGD and non-RGD binding sites for multiple integrins. Adjacent to the RGD motif is the sequence SLAYGLR (SVVYGLR in human) which serves as a cryptic binding site for additional integrins: it is masked in full length OPN but is exposed following OPN cleavage by multiple proteases in tumors and sites of tissue injury.
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Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit
R&D Systems | Catalog # MOST00
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Urine (50 µL)
Sensitivity
8.5 pg/mL
Assay Range
39-2500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
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Product Summary for Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit
The Quantikine Mouse OPN Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse OPN in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse OPN and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse OPN showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse OPN.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Mouse, Rat
Specificity
Natural and recombinant mouse and rat OPN. This assay cross-reacts 8.2% with recombinant Human Osteopontin.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 62 | 67 | 59 |
| Mean (pg/mL) | 96.0 | 236 | 1085 | 116 | 279 | 1076 |
| Standard Deviation | 7.3 | 13.8 | 43.4 | 9.8 | 22.5 | 90.0 |
| CV% | 7.6 | 5.8 | 4.0 | 8.4 | 8.1 | 8.4 |
Recovery for Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit
The recovery of mouse OPN spiked to three levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Mouse Cell Culture Supernates (n=7) | 98 | 87-106 |
| Rat Cell Culture Supernates (n=7) | 93 | 82-107 |
Linearity
To assess the linearity of the assay, samples containing mouse or rat OPN were diluted with Calibrator Diluent and then assayed.
Scientific Data Images for Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit
Mouse/Rat Osteopontin/OPN ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Osteopontin/OPN
Long Name
Secreted Phosphoprotein 1 [BNSP]
Alternate Names
Eta-1, OPN, Spp1
Gene Symbol
SPP1
Additional Osteopontin/OPN Products
Product Documents for Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Citations for Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit
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Protocols
View specific protocols for Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit (MOST00):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
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- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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