Human Osteopontin(OPN) enzyme-linked immunoassay kit

CAT: EH0057 Datasheet
Specification 96 Test
Sensitivity 0.04 ng/ml (50 μl);0.12 ng/ml (10 μl)
Standard Curve Range 0.31~20 ng/ml
Standard Curve Gradient 7 Points
Number of Incubations 2
Sample Volume 50 μl/10 μl
Type Ready-to-Use
Operation Duration 120min
ng/ml O.D. Average Corrected
0.00 0.0165 0.0176 0.0171
0.31 0.0363 0.0396 0.0380 0.0209
0.63 0.0749 0.0860 0.0805 0.0634
1.25 0.1778 0.1955 0.1867 0.1696
2.50 0.4225 0.4620 0.4423 0.4252
5.00 1.0110 1.0540 1.0325 1.0155
10.00 2.0350 2.1320 2.0835 2.0665
20.00 3.5040 3.4120 3.4580 3.4410

Precision

Intra-assay Precision Inter-assay Precision
Sample Number S1 S2 S3 S1 S2 S3
22 22 22 6 6 6
Average(ng/ml) 0.42 2.09 6.22 0.51 2.10 6.03
Standard Deviation 0.02 0.09 0.48 0.02 0.10 0.37
Coefficient of Variation(%) 4.9 4.5 7.7 4.3 4.7 6.1

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of human OPN into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 83% to 117% with an overall mean recovery of 108%.

Sample Values

Sample Matrix Sample Evaluated Range (ng/ml) Detectable (%) Mean of Detectable (ng/ml)
Serum 30 1.1-6.74 100 3.97

Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of OPN in this assay. No medical histories were available for the donors.

Background: OPN

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 ad¬hesion, and migration. Its expression is upregulat¬ed 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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