Components
Pre-coated 96 well plate
Standard
Assay Diluent concentrate
Biotinylated Detection Antibody
SAV-HRP
Wash Buffer
Chromogen
Stop Solution
Adhesive Plate Covers
Material not included
Microplate reader capable of reading at a wavelength of 450 nm
Variable volume Microdrop Pipettes
Adjustable multi-channel pipette
Reagent reservoir
Bottle or dishwashing systems
Distilled or deionized water
Serological pipettes
Disposable straw Tips
Sample matrix free of endogenous interferon
Assay Time
Less than 48 hours
Reagent Preparation
1. The buffer concentrate should be brought to room temperature and diluted before starting the test procedure.
2. If crystals form in the buffer concentrate, heat gently until the crystals are completely dissolved. See the kit manual for the rest.
Assay Procedure
1. Add Incubation Buffer to all wells except the chromogen blank.
2. Add the standard, buffer solution, or cell culture medium sample to the appropriate wells. for
Serum and control samples, add standard dilution buffer, then add samples to
suitable well. Leave the chromogen blank holes blank.
3. Tap the side of the plate to mix. Cover the plate with a plate lid and incubate at room temperature for 2-3 hours.
4. Aspirate the solution thoroughly and wash the wells 4 times with wash buffer.
5. Add Biotin Conjugate solution to each well except the chromogen blank.
6. Cover the plate and incubate at room temperature for 1-2 hours.
7. Aspirate the solution thoroughly and wash the wells 4 times with wash buffer.
8. Add Streptavidin-HRP solution to each well except the chromogen blank.
9. Cover the plate with a plate cover and incubate at room temperature.
10. Aspirate the wells thoroughly and wash the wells 4-6 times with wash buffer.
11. Add stable chromogen to each well. The substrate solution begins to turn blue.
12. Incubate at room temperature for 30-45 minutes in the dark.
13. Add stop solution to each well. Tap the side of the plate to mix. The solution in the well changed from blue to yellow.
Calculation of Results
The titer of interferon in the sample can be determined by plotting the optical density (OD) by fitting a 4-parameter standard curve. Standard and sample ozone depletion should be subtracted from blank ozone depletion to eliminate background. A typical standard curve for this analysis is shown below. This example is for illustration only and should not be used to calculate unknowns.
Assay Precision
12.5-400 pg/ml
Format
Sandwich ELISA Kit
Precaution of Use
All products are supplied for research and laboratory use only.
Handling Advice
All blood components and biological materials should be treated with precautions as potentially hazardous. Strictly follow the management principles of the Centers for Disease Control and Prevention, Occupational Safety and Health Administration for handling and disposing of infectious agents.
Storage
The ELISA Kits are shipped at 2 to 8°C. Upon receipt, store the kits at 2 to 8°C in dark.
Expiry Date
Stability If properly stored, all components are stable for up to 12 months. For expiry dates for the entire kit, see the kit label. The expiration date of each ingredient is shown on the bottle label. The expiration date of a kit ingredient is guaranteed only if the ingredient is properly stored and, in the event of repeated use of an component, the reagent will not be contaminated by the first treatment.
Note
Each production lot of this ELISA kit is quality tested to meet criteria such as sensitivity, specificity, precision and lot-to-lot consistency. See the manual for more information on validation.
