Osteopontin ELISA Kit

Catalog No. ABIN411344

This Colorimetric ELISA kit is designed for the quantitative measurement of Mouse Osteopontin. There are 2 publications available.

Quick Overview for Osteopontin ELISA Kit (ABIN411344)

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

Binding Specificity: AA 17-294

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156 pg/mL - 10000 pg/mL

Application: ELISA

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

Product Details Osteopontin ELISA Kit

Minimum Detection Limit: 156 pg/mL

Purpose: Sandwich Mouse OPN / Osteopontin ELISA Kit to quantitate Mouse Spp1 in cell culture supernatants, serum, plasma (heparin, EDTA) and urine.

Analytical Method: Quantitative

Specificity: Expression system for standard: NS0
Immunogen sequence: L17-N294

Capture antibody and Detection antibody: monoclonal antibody from rat, polyclonal antibody from goat

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

Sensitivity: < 10 pg/mL

Components:

1. Pre-coated 96-well strip microplate
2. Standard
3. Biotinylated antibody (100x)
4. Avidin-Biotin-Peroxidase Complex (100x)
5. Sample Diluent
6. Antibody Diluent
7. Avidin-Biotin-Peroxidase Diluent
8. Color Developing Reagent (TMB)
9. Stop Solution
10. Wash Buffer (25x)
11. Adhesive plate sealers

Material not included: 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. 500 mL graduated cylinders. Test tubes for dilution.

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.

Sample Volume: 100 μL

Plate: Pre-coated

Protocol: ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection antibody specific for target 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 target amount in sample captured in plate.

Assay Precision: Intra-Assay Precision (CV%): 4.8%, 7.5%, 7.5%
Inter-Assay Precision (CV%): 4.9%, 7.1%, 9%

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 (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Expiry Date: 12 months

References for Osteopontin ELISA Kit (ABIN411344)

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).

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:

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 mouse OPN, consisting of 284 amino acid sequence with the molecular mass of 31.5KDa. As a result of glycosylation, the molecular mass is 65KDa.

Gene Full Name: secreted phosphoprotein 1

Gene ID: 20750

UniProt: P10923

Pathways: Regulation of Cell Size