Human OPN / Osteopontin ELISA Kit PicoKine®

Osteopontin/OPN ELISA kit for Human

Human OPN / Osteopontin ELISA Kit PicoKine™ (96 Tests). Quantitate Human SPP1 in cell culture supernatants, serum, plasma (heparin, EDTA) , human milk and urine. Sensitivity: 10pg/ml. Cited in 10 publication(s).

Product Info Summary

SKU: EK0482
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, serum, plasma (heparin, EDTA) , human milk and urine.

Product Name

Human OPN / Osteopontin ELISA Kit PicoKine®

View all Osteopontin/OPN ELISA kits

SKU/Catalog Number

EK0482

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

Human OPN / Osteopontin ELISA Kit PicoKine™ (96 Tests). Quantitate Human SPP1 in cell culture supernatants, serum, plasma (heparin, EDTA) , human milk and urine. Sensitivity: 10pg/ml.

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cite This Product

Human OPN / Osteopontin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0482)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: I17-N300

Sensitivity

<10 pg/ml

Assay Range

156 pg/ml - 10,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK0482 for more details

Cross-reactivity

There is no detectable cross-reactivity with other relevant proteins.

Reactive Species

EK0482 is reactive to SPP1 in Human samples

Validated Sample Types

cell culture supernatants, serum, plasma (heparin, EDTA) , human milk and urine.

Application Guarantee

EK0482 is guaranteed for ELISA in Human by Boster Guarantee

See how Boster Bio validate our ELISA kits: ELISA Validation Information

Background of Osteopontin/OPN

Osteopontin (OPN) also called urinary stone protein, secreted phosphoprotein 1 (SPP1), bone sialoprotein, and early T lymphocyte activation 1 (ETA1). Osteopontin is a phosphorylated glycoprotein secreted to the mineralizing extrOPNllular matrix by osteoblasts during bone development. It is believed to facilitate the attachment of osteoblasts and osteoclasts to the extrOPNllular matrix, allowing them to perform their respective functions during osteogenesis. Osteopontin is presumably involved in stone formation as stone matrix. The deduced protein sequence reveals a 317-amino acid protein (34,982 Da) containing a 16-amino acid hydrophobic signal sequence and a 33,352-Da protein destines to undergo extensive post-translational modifications before being secreted from the cell. The gene is located on human chromosome 4. The standard product used in this kit is recombinant human OPN, consisting of 17-314 amino acid sequence with the molecular mass of 32.9KDa. As a result of glycosylation, the molecular mass is 60-65KDa.

Kit Components

Catalog Number Description Quantity
EK0482-CAP Anti-Human SPP1 Pre-coated 96-well strip microplate 1
EK0482-ST Human SPP1 Standard 2 vials, 10 ng/tube
EK0482-DA Human SPP1 Biotinylated antibody (100x) 100ul
AR1103 Avidin-Biotin-Peroxidase Complex (100x) 100ul
AR1106-1 Sample Diluent 30ml
AR1106-2 Antibody Diluent 12ml
AR1106-3 Avidin-Biotin-Peroxidase Diluent 12ml
AR1104 Color Developing Reagent (TMB) 10ml
AR1105 Stop Solution 10ml
AR1106-5 Wash Buffer (25x) 20ml
PLA-SEA Adhesive plate sealers 4

*The kit components are not available for individual purchase.

Materials Required But Not Included In Kit

  • Microplate Reader capable of reading absorbance at 450nm.
  • Incubator.
  • Automated plate washer (optional).
  • Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
  • Multichannel pipettes are recommended for large amount of samples.
  • Deionized or distilled water.
  • 500ml graduated cylinders.
  • Test tubes for dilution.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)015631262512502500500010000
O.D.0.0560.1100.1560.2440.4570.9461.6772.085

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect Osteopontin/OPN, Dilution ratio of 1:100, concentration in plasma is 39-234 ng/ml..

Some articles we found to cite concentrations of Osteopontin/OPN in samples: 24338767 (Pubmed IDs).

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human OPN / Osteopontin ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)3051771498531416115408
Standard deviation14.03138.13304.0816.64133.71383.96
CV (%)4.6%7.8%6.1%5.3%8.3%7.1%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of Osteopontin/OPN in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 130530927232230218.426%
Sample 21771175117091739174222.461.2%
Sample 349854178447644054511294.966.5%
*number of samples for each test n=16.

Gene/Protein Information For SPP1 (Source: Uniprot.Org, NCBI)

Gene Name

SPP1

Full Name

Osteopontin

Weight

35.423kDa

Superfamily

osteopontin family

Alternative Names

BNSP; Bone sialoprotein 1; Eta-1; MGC110940; Nephropontin; OPN; Osteopontin; secreted phosphoprotein 1bone sialoprotein I, early T-lymphocyteactivation 1); secreted phosphoprotein-1 (osteopontin, bone sialoprotein); Spp1; SPP-1; SPP1/CALPHA1 fusion; Urinary stone protein; uropontin SPP1 BNSP, BSPI, ETA-1, OPN secreted phosphoprotein 1 osteopontin|SPP1/CALPHA1 fusion|early T-lymphocyte activation 1|nephropontin|osteopontin/immunoglobulin alpha 1 heavy chain constant region fusion protein|secreted phosphoprotein 1 (osteopontin, bone sialoprotein I, early T-lymphocyte activation 1)|secreted phosphoprotein 1 variant 6|urinary stone protein|uropontin

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

For more info on SPP1, check out the SPP1 Infographic

SPP1 infographic

We have 30,000+ of these available, one for each gene! check them out.

In this infographic you will see the following information for SPP1: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].

Hello CJ!

EK0482 has been cited in 10 publications:

*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.

Renoprotective effects of berberine as adjuvant therapy for hypertensive patients with type 2 diabetes mellitus: Evaluation via biochemical markers and color Doppler ultrasonography

The association between serum angiogenin and osteopontin levels and coronary collateral circulation in patients with chronic total occlusion

Predictive Value of Serum Bone Sialoprotein in Patients with Bone Metastasis of Non-Small Cell Lung Cancer

Diminished membrane recruitment of Akt is instrumental in alcohol‐associated osteopenia via the PTEN/Akt/GSK‐3β/β‐catenin axis

Expression of NF-κB and osteopontin of synovial fluid of patients with knee osteoarthritis

Combination of nanolamellae and PDA coating on promoting the long-term adhesion, proliferation, and differentiation of osteoblasts

Hypoxia‐driven paracrine osteopontin/integrin αvβ3 signaling promotes pancreatic cancer cell epithelial–mesenchymal transition and cancer stem cell‐like properties by modulating forkhead box protein M1

Hypoxia‐driven paracrine osteopontin/integrin αvβ3 signaling promotes pancreatic cancer cell epithelial–mesenchymal transition and cancer stem cell‐like properties by modulating forkhead box protein M1

The interaction of MMP-2/B7-H3 in human osteoporosis

Hypoxia-driven paracrine osteopontin/integrin αvβ3 signaling promotes pancreatic cancer cell epithelial-mesenchymal transition and cancer stem cell-like properties by modulating forkhead box protein M1

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8 Customer Q&As for Human OPN / Osteopontin ELISA Kit PicoKine®

Question

In this part of the EK0482 protocol, what should 0.2?m filter mean? Is it mm? "Centrifuge for 15 min at 1500 x g at 2-8°C. Collect the aqueous layer and repeat this process 3 times. Filter through a 0.2?m filter. Assay immediately or aliquot and store at -80°C.''

Verified customer

Asked: 2021-10-31

Answer

It should be "Filter through a 0.2μm filter" in the Human OPN / Osteopontin ELISA Kit PicoKine® (EK0482) protocol.

Boster Scientific Support

Answered: 2021-11-01

Question

Q: how are cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2020-04-17

Answer

A: for those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present in the product insert. Components in lysate and lysis buffer may impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2020-04-17

Question

Q: Can Osteopontin ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

Verified Customer

Verified customer

Asked: 2019-08-29

Answer

A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not valid for other sample types than we have tested: it means further investigation is required. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been evaluated by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please see the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.

Boster Scientific Support

Answered: 2019-08-29

Question

Can I use this for detection in both sputum and serum samples? Keyword: applications

Verified Customer

Verified customer

Asked: 2019-06-28

Answer

This product is for quantitative detection of human OPN in cell culture supernates, serum, plasma(heparin, EDTA), human milk and urine. We did not do a related test about sputum, we suggest you to do a trial test first.

Boster Scientific Support

Answered: 2019-06-28

Question

Q: can you suggest the dilution ratio of serum samples for detection of Osteopontin in Human urine? I am trying to measure a a number of analytes and it requires 100ul of diluted samples for each well. We have limited sample volumes so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2018-12-26

Answer

A: unable to know the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the Osteopontin ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2018-12-26

Question

Q: is it okay to use citrate plasma as samples in Human Osteopontin Picokine® ELISA Kit (Catalog # EK0482)?

A. Chakrabarti

Verified customer

Asked: 2018-09-13

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of Osteopontin causing disruption of its protein structure. Osteopontin may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes needed for detection and block the antigen antibody reaction. We have tested the Osteopontin ELISA, treating samples with a number of anticoagulants and decided that heparin or EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2018-09-13

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

P. Davis

Verified customer

Asked: 2018-02-06

Answer

A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions

Boster Scientific Support

Answered: 2018-02-06

Question

Q: how much samples can be assayed in a Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2018-02-01

Answer

A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may vary slightly by kit so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2018-02-01

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