Staphylococcal aureus Enterotoxin Total (SET) ELISA Kit
For reference only. Please follow the manual included in your kit for instructions.
Catalog Number
RDF-E118
The Staphylococcal aureus Enterotoxin Total (SET) ELISA Kit (RDF-E118) is an enzyme-linked immunosorbent assay for quantitative detection of SET in Food samples with a standard range of . This kit uses the Traditional ELISA format for colorimetric detection of natural SET in samples including Milk,Yogurt, Milk powder. The kit is easy to use and comes with a pre-coated microplate and all necessary reagents and buffers. The standard curve generated in the experiment is used to calculate the concentration of SET in samples. For research use only.
Product Specifications
Product Name
Staphylococcal aureus Enterotoxin Total (SET) ELISA Kit
Catalog Number
RDF-E118
Species Reactivity
Food
Species
Food
Assay Length
120 minutes
Experimental Method
Sandwich
Detection Method
Colorimetric
Sample Type
Milk,Yogurt, Milk powder
Storage
2-8°C,Avoid freeze
Shelf Life
12 monthes
Product Details
Intended Use
For the qualitative detection of Staphylococcal aureus Enterotoxin Total (SET) concentration in milk, milk powder, and yogurt samples.
Assay Principle
This kit uses Sandwich-ELISA as the method for the qualitative detection. It can detect staphylococcal aureus enterotoxin (SET) (A, B, C, D and E) in samples, such as, milk, milk powder, and yogurt. This kit is composed of ELISA Microtiter plate, HRP conjugate, antibody working solution, control and other supplementary reagents. The microtiter plate in this kit has been pre-coated with coupled antibodies. The sample is added to the wells of the ELISA microtiter plate, and the SET in the sample is combined with the pre-coated antibody to form an SET-antibody compound. Free components are washed away. The Antibody Working Solution and HRP Conjugate are added to each well and react with the compound to form “antibody-SET-HRP conjugate” compound. The substrate reagent is added to initiate the color developing reaction. The presence of SET can be determined according to the OD value by using a microplate reader with 450 nm (630 nm) wavelength.
Application Data
Reagents and Materials Provided
| Reagent | Quantity |
|---|---|
| ELISA Microtiter plate | 96 wells |
| Antibody Working Solution | 12 mL |
| HRP Conjugate | 12 mL |
| Positive Control | 2 mL |
| Negative Control | 2 mL |
| Substrate Reagent A | 6 mL |
| Substrate Reagent B | 6 mL |
| Stop Solution | 6 mL |
| Acidity regulators | 10 mL |
| 10×Concentrated Wash Buffer | 2*40 mL |
| Plate Sealer | 3 |
| Sealed Bag | 1 |
| Manual | 1 |
All reagent bottle caps should be tightly closed to prevent evaporation and microbial contamination.
Materials Required but not Supplied
- Equipment: Microplate reader, Printer, Homogenizer, Oscillators, Centrifuge, Graduated pipette, Balance (sensibility 0.01 g).
- High-precision Micropipettes: single-channel 20-200 µL, 100-1000 µL.
Storage
- Store the kit at 2-8℃. Do not freeze any kit components.
- Return any unused microwells to their original foil bag and reseal them together with the desiccant provided. Continue to store at 2-8℃.
- Shelf life is approximately 12 months from date of manufacture. Check packaging for expiry date.
Sample Preparation
Milk:
- Take 10 mL of milk samples into 50 mL centrifuge tube, centrifuge at 10000 r/min for 10 min at 4-10℃ (If a refrigerated centrifuge is not available, chill sample to approx 4-10℃ prior to centrifugation).
- Take 100 μL of middle layer for analysis.
- Yogurt:
- Weigh 1±0.05 g of yogurt samples into 50 mL centrifuge tube, add 5 mL of deionized water. Oscillate fully for 5 min.
- Centrifuge at 10000 r/min for 10 min at 4-10℃ (If a refrigerated centrifuge is not available, chill sample to approx 4-10℃ prior to centrifugation).
- Take all middle layer to another centrifuge tube, add Acidity regulators and adjust the pH to 7.0.
- Take 100 μL for analysis.
- Milk powder:
- Weigh 1±0.05 g of milk powder samples into 50 mL centrifuge tube, add 7 mL of deionized water. Oscillate fully for 2 min.
- Take 100 μL for analysis.
Reagent Preparation
Solution preparation:
Solution 1: Wash Buffer
Dilute the 10×Concentrated Wash Buffer with deionized water. (10×Concentrated Wash Buffer (V): Deionized water (V) =1:9).
Notes:- Bring all reagents and samples to room temperature before use.
- Turn on the microplate reader 30 min in advance to preheat the instrument and set the testing parameters.
- All experimental apparatus should be clean, and disposable pipette should be used and changed frequently to avoid cross-contamination during the experiment.
Assay Procedure
Bring all reagents and samples to room temperature (25℃) before use. All reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Any unused Microtiter plate wells should be resealed as soon as possible and stored at 2-8℃.
- Number: number the samples and standards in order (multiple wells), and keep a record of the plate layout. Postive and Negative controls and Samples must be tested in duplicate.
- Add Sample: add 100 μL of Postive/Negative Control or Sample to each well, and cover the plate with plate sealer. Oscillate gently for 5 s to mix thoroughly, then incubate at 37℃ for 60 min in the dark.
- Wash: Carefully remove the plate sealer, then remove the liquid from each well. Immediately add 250 μL of Wash Buffer (Solution 1) to each well and wash. Repeat wash procedure 5 times, 10 s intervals each time. Invert the plate and pat it against a thick sheet of absorbent paper (If there are bubbles in the wells, clean pipette tips can be used to pop them).
- Antibody Working Solution: Add 100 uL Antibody Working Solution to each well, and cover the plate with plate sealer. Oscillate for 5 s gently to mix thoroughly. Incubate at 37℃ for 30 min in the dark.
- Wash: repeat step 3.
- HRP Conjugate: add 100 μL of HRP Conjugate to each well, incubate at 37℃ for 15 min in the dark.
- Wash: repeat step 3.
- Color Development: add 50 μL of Substrate Reagent A to each well, and then add 50 μL of Substrate Reagent B. Gently oscillate for 5 s to mix thoroughly. Incubate at 37℃ for 15 min in the dark (The reaction time can be extended according to the amount of color change).
- Stop Reaction: add 50 μL of Stop Solution to each well. Gently oscillate to mix thoroughly.
- OD Measurement: determine the optical density (OD value) of each well at 450 nm (reference wavelength 630 nm) with a microplate reader. This step should be performed no later than 5 min after the reaction has been stopped.
- REFERENCE VALUE
- Normally, Average absorbance of negative control < 0.2 and Average absorbance of positive control > 0.5.
- INTERPRETATION OF THE RESULTS
- Cut Off = 0.2 + Average absorbance of negative control
- 1. Positive result: Average absorbance of sample ≥ Cut Off.
- 2. Negative result: Average absorbance of sample < Cut Off.
Calculation of Results
Product Data
Reaction conditions (Incubation time and temperature)
25℃; 60 min, 30 min, 15 min, 15 min