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
A microtiter plate reader capable of measuring at or near 450 nm.
Calibrate adjustable precision pipettes, preferably disposable
plastic head.
Distilled or deionized water.
Dishwasher: automatic or manual (spray bottle, manifold)
dispenser, etc.).
Data analysis and graphing software.
Dilution and mixing standards in glass or plastic tubes.
An absorbent paper towel.
Various sizes of calibration beakers and graduated cylinders.
Assay Time
Less than 4 hours
Reagent Preparation
1. Negative and positive controls
Reconstitute the lyophilized Controls to the volume specified on the vial label with distilled water.
2. HRP Conjugate Working Solution
Depending on the number of wells to be used, dilute the concentrated conjugate with the conjugate buffer in a clean glass vial.
3. Working Washing Solution
Dilute Super-Wash with distilled water.
Assay Procedure
Repeat determination of samples is recommended.
1. Select the number of entries required for running.
2. Secure the strip to the securing frame.
3. Move qc products and samples into appropriate holes.
4. Move incubation buffer into each well.
5. Incubate the board on a horizontal shaker at room temperature for 1 hour.
6. Suck out the contents of each well.
7. Wash the plate 3-5 times with working washing solution.
8. Remove any remaining droplets by tapping the board on blotting paper.
9. Move the working coupling solution into each well.
10. Incubate the board on a horizontal shaker at room temperature for 1 hour.
11. Suck out the contents of each well.
12. Wash the plate 3-5 times with working detergent.
13. Remove remaining droplets by tapping the board on blotting paper.
14. Distribute the chromogen (TMB) solution to each well.
15. Incubate the board on a horizontal shaker at room temperature for 15 minutes.
16. Add stop solution to each well to stop color development.
17. Read the plate at 450 nm against any reference filter of 630 to 750 nm. Record the absorbance value.
Calculation of Results
1. Read the absorbance at 450nm. Read the plate within 2 hours of adding the stop solution.
2. Generate a standard curve using curve fitting software.
3. Read the concentrations of unknown samples and controls from the standard curve. The sample dilution is corrected by an appropriate factor, resulting in a number of values (s) for the sample.
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.
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.
Datasheet