Osteopontin ELISA Kit

Catalog No. ABIN411343

Human Osteopontin ELISA Kit Colorimetric assay for quantification of Human Osteopontin and has been mentioned in 4+ publications.

Quick Overview for Osteopontin ELISA Kit (ABIN411343)

Target: Osteopontin (SPP1) (Secreted phosphoprotein 1 (SPP1))

Binding Specificity: AA 17-300

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156-10000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA), Milk, Urine

Product Details Osteopontin ELISA Kit

Minimum Detection Limit: 156 pg/mL

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human OPN

Brand: PicoKine™

Analytical Method: Quantitative

Specificity: Expression system for standard: NSO
Immunogen sequence: I17-N300

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: NSO
Immunogen sequence: I17-N300

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Belongs to the osteopontin family.

Tissue Specificity: Bone. Found in plasma.

Plate: Pre-coated

Protocol: human OPN ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for OPN has been precoated onto 96-well plates. Standards(NSO, I17-N300) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for OPN is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human OPN amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 10000pg/mL, 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312pg/mL, 156pg/mL human OPN standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of human cell culture supernatants, serum, plasma(heparin, EDTA), milk or urine to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each human OPN standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(ng/ml): 2.5, Standard deviation: 0.105, CV(%): 4.2
• Sample 2: n=16, Mean(ng/ml): 4.3, Standard deviation: 0.146, CV(%): 3.4
• Sample 3: n=16, Mean(ng/ml): 6.1, Standard deviation: 0.317, CV(%): 5.2,
• Sample 1: n=24, Mean(ng/ml): 2.6, Standard deviation: 0.161, CV(%): 6.2
• Sample 2: n=24, Mean(ng/ml): 3.9, Standard deviation: 0.226, CV(%): 5.8
• Sample 3: n=24, Mean(ng/ml): 6.3, Standard deviation: 0.353, CV(%): 5.6

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

References for Osteopontin ELISA Kit (ABIN411343)

Qin, Yan, Wang, Xu, Wang, Zhu, Shi, Li, Zhang, Chen: "Cancer-associated Fibroblast-derived IL-6 Promotes Head and Neck Cancer Progression via the Osteopontin-NF-kappa B Signaling Pathway." in: Theranostics, Vol. 8, Issue 4, pp. 921-940, (2018) (PubMed).

El-Din, Amin, Egiza: "Utility of Tissue Inhibitor Metalloproteinase-1 and Osteopontin as Prospective Biomarkers of Early Cardiovascular Complications in Type 2 Diabetes." in: Open access Macedonian journal of medical sciences, Vol. 6, Issue 2, pp. 314-319, (2018) (PubMed).

Sun, Tu, Chen, Yuan, Liu, Dong, Yuan: "Three-dimensional combined biomarkers assay could improve diagnostic accuracy for gastric cancer." in: Scientific reports, Vol. 7, Issue 1, pp. 11621, (2017) (PubMed).

Chen, Sun, He, Gong, Xu, Yuan: "Serum OPN expression for identification of gastric cancer and atrophic gastritis and its influencing factors." in: PLoS ONE, Vol. 9, Issue 12, pp. e114005, (2014) (PubMed).

Target Details for Osteopontin

Target: Osteopontin (SPP1) (Secreted phosphoprotein 1 (SPP1))

Alternative Name: SPP1

Background:

Protein Function: Binds tightly to hydroxyapatite. Appears to form an integral part of the mineralized matrix. Probably important to cell-matrix interaction.

Background: 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.

Synonyms: Osteopontin,Bone sialoprotein 1,Nephropontin,Secreted phosphoprotein 1,SPP-1,Urinary stone protein,Uropontin,SPP1,BNSP, OPN,PSEC0156,

Full Gene Name: Osteopontin

Cellular Localisation: Secreted.

Gene ID: 6696

UniProt: P10451

Pathways: Regulation of Cell Size