Bovine Osteopontin ELISA Kit

Sample

Serum, plasma, tissue homogenates and other biological fluids

Species Reactivity

Bovine

Detection Method

sELISA

Intended Use

This kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of OPN in bovine serum, plasma, tissue homogenates and other biological fluids.

Contents of Kit

1. Pre-coated, ready to use 96-well strip plate
2. Standard
3. Detection Solution A
4. Detection Solution B
5. Wash Buffer 30×
6. Standard Diluent
7. TMB Substrate
8. Stop Solution
9. Instruction manual 1
10. Plate sealer for 96 wells 2

Storage

1. For unopened kits: All the reagents should be stored at -20°C upon receipt.
2. For opened kits: Once the kit is opened, the remaining reagents still need to be stored according to the above storage conditions. In addition, return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
Note:
For the expiration date of the kit, please refer to the label on the kit box. All components are stable until this date.
It is highly recommended to use the remaining reagents within 1 month of opening.

Performance Characteristics

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage conditions.
Note:
To minimize unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity, and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level OPN were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level OPN were tested on 3 different plates, 8 replicates in each plate.
CV (%) = SD/mean × 100
Intra-Assay: CV<10% Inter-Assay: CV<12%

Detection Range

0.156-10 ng/mL. The concentrations used for creating the standard curve were 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL.

Sensitivity

The minimum detectable dose of OPN is typically less than 0.069 ng/mL. The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.

Standard Curve

Average the duplicate readings for each standard, control and sample, then subtract the average zero standard optical density. Construct a standard curve by plotting the mean O.D. and concentration for each standard and draw a best fit curve through the points on the graph or create a standard curve on log-log graph paper with OPN concentration on the y-axis and absorbance on the x-axis. Using plotting software, (for instance, curve expert 1.30), is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.