Osteopontin ELISA Kit

Catalog No. ABIN1112773

Product Details Osteopontin ELISA Kit

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

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 156-10000 pg/mL

Minimum Detection Limit: 156 pg/mL

Application: ELISA

Analytical Method: Quantitative

Sensitivity: < 10 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Mouse OPN antibody 2. Lyophilized Mouse OPN standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-Mouse OPN antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-OPN polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-OPN polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the OPN amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of OPN can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Tissue lysate, body fluids and cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 2000 × g for 20 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with heparin, citrate or EDTA as the anticoagulant. Centrifuge for 20 min at 2000 x g within 30 min of collection. Analyze immediately or aliquot and store frozen at -20 °C. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Target Details for Osteopontin

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

Alternative Name: OPN (SPP1 Products)

Synonyms: BNSP ELISA Kit, BSPI ELISA Kit, ETA-1 ELISA Kit, OPN ELISA Kit, 2AR ELISA Kit, Apl-1 ELISA Kit, Bsp ELISA Kit, Eta ELISA Kit, OP ELISA Kit, Opn ELISA Kit, Opnl ELISA Kit, Ric ELISA Kit, Spp-1 ELISA Kit, OSP ELISA Kit, zgc:111821 ELISA Kit, secreted phosphoprotein 1 ELISA Kit, SPP1 ELISA Kit, Spp1 ELISA Kit, spp1 ELISA Kit

Background: Osteopontin (OPN), also known as bone sialoprotein I (BSP-1 or BNSP), secreted phosphoprotein 1 (SPP1), is a human gene product. The gene has 7 exons, spans 5 kilobases in length and in humans it is located on 4q13. The protein is composed of ~300 amino acids residues and has ~30 carbohydrate residues attached including 10 sialic acid residues, which are attached to the protein during post-translational modification in the Golgi apparatus. Osteopontin has been implicated as an important factor in bone remodeling. Specifically, research suggests it plays a role in anchoring osteoclasts to the mineral matrix of bones.

Pathways: Regulation of Cell Size