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
Precision pipette, disposable 5-1000µL plastic tip, 5-15m plastic pipette
A two-liter glass or plastic container to prepare wash buffer
Automatic 96-well dish washer
1.5mL polypropylene or polyethylene tube prep standard
Disposable reagent reservoir
15mL plastic tube to prepare streptavidin-hrp solution
Standard ELISA reader for absorbance measurements at 450nm and 550nm.
Graphics paper or computerized curve fitting statistical software package
Assay Time
Less than 4 hours
Reagent Preparation
1. Wash Buffer
Wash buffers must be at room temperature before use in assays. If it becomes apparent, do not wash with water that has been contaminated during storage.
When using partial plates, store the reconstituted wash buffer at 2-8°C.
2. Standards
The lyophilized standard was reconstituted and used, per vial. Prepare standards before use and use within one hour of reconstitution. Do not store refactored criteria.
Assay Procedure
1. Sample Incubation
2. Plate Washing
3. Biotinylated Antibody Reagent Incubation
4. Streptavidin-HRP Solution Preparation and Incubation
5. Substrate Incubation and Stop Step
6. Absorbance Measurement
Note: Evaluate the plate within 30-45 minutes of stopping the reaction.
Calculation of Results
1.Calculate the mean absorbance values for each set of replicate standards and samples.
2. Create a standard curve by plotting the mean absorbance of each standard concentration on the ordinate and the Rat IFN gamma concentration on the abscissa.
3. To determine the circulating Rat IFN gamma concentration for each sample, first find the mean absorbance value on the ordinate and extend a horizontal line onto the standard curve.
4. At the intersection, the vertical line is extended to the abscissa and the corresponding Rat IFN gamma concentration is read.
5. The sample has been diluted, so the concentration read from the standard curve must be multiplied by the dilution factor.
Assay Precision
8-500 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.
