Components
Pre-coated, ready to use 96-well strip plate
Plate sealer for 96 wells
Standard
Standard Diluent
Detection Reagent A
Detection Reagent B
Assay Diluent A
Assay Diluent B
TMB Substrate
Stop Solution
Wash Buffer (30 x concentrate)
Instruction manual
Material not included
1.Microplate reader with 450 ± 10nm filter.
2.Precision single or multi-channel pipettes and disposable tips.
3.Tubes for diluting samples.
4.Deionized or distilled water.
5.Absorbent paper for blotting the microtiter plate.
6.Container for Wash Solution.
7.0.01Mol/L (or 1x) Phosphate Buffered Saline(PBS), pH 7.0-7.2.
Assay Procedure
The microtiter plate provided in this kit has been pre-coated with an antibody specific to the index. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to the index. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain the index, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm.
Assay Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%
Precaution of Use
The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
Handling Advice
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.
Storage Comment
The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20°C upon being received. After receiving the kit , Substrate should be always stored at 4°C.Other reagents are kept according to the labels on vials. But for long term storage, please keep the whole kit at -20°C. The unused strips should be kept in a sealed bag with the desiccant provided to minimize exposure to damp air. The test kit may be used throughout the expiration date of the kit (six months from the date of manufacture). Opened test kits will remain stable until the expiring date shown, provided it is stored as prescribed above.
Note
The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20°C upon being received. After receiving the kit , Substrate should be always stored at 4°C.Other reagents are kept according to the labels on vials. But for long term storage, please keep the whole kit at -20°C. The unused strips should be kept in a sealed bag with the desiccant provided to minimize exposure to damp air. The test kit may be used throughout the expiration date of the kit (six months from the date of manufacture). Opened test kits will remain stable until the expiring date shown, provided it is stored as prescribed above.
Restrictions
For Research Use only
