Components
Assay plate (12 × 8 coated Microwells)
Standard (freeze dried)
Biotin-antibody (100 × concentrate)
HRP-avidin (100 × concentrate)
Biotin-antibody Diluent
HRP-avidin Diluent
Sample Diluent
Wash Buffer (25 × concentrate)
TMB Substrate
Stop Solution
Adhesive Strip (for 96 wells)
Instruction manual
Material not included
Microplate reader capable of measuring absorbance at 450nm, with the correction wavelength set at 540nm or 570nm.
An incubator which can provide stable incubation conditions up to 37°C ± 0.5°C.
Squirt bottle, manifold dispenser or automated microplate washer.
Absorbent paper for blotting the microtiter plate.
100mL and 500mL graduated cylinders.
Deionized or distilled water.
Pipettes and pipette tips.
Test tubes for dilution.
Calculation of Results
Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit.
Assay Precision
Intra-assay precision (precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess precision.
Inter-assay precision (precision between assays): Three samples of known concentration were tested in twenty assays to assess precision.
Intra-assay: CV% less than 8%
Inter-assay: CV% less than 10%
Precaution of Use
The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face and clothing protection when using this material.
Handling Advice
1.All reagents must be at room tempecanineure (18 °C to 25 °C) before running assay.
2.Do not expose kit reagents to strong light during storage or incubation.
3.Improper or insufficient washing at any stage of the procedure will result in either false positive or false negative results.
4.Avoid contact of stop solution with skin or eyes. If contact occurs, immediately flush area with copious amounts of water.
5.Do not use TMB substcaninee solution if it has begun to turn blue.
6.Do not expose bleach to work area during actual test procedure because of potential interference with enzyme activity.
Storage Comment
May be stored at 2-8°C for up to 1 month. For long term storage, please store at -20°C. Try to keep assay plate in a sealed aluminium foil bag and avoid dampness.
Note
May be stored at 2-8°C for up to 1 month. For long term storage, please store at -20°C. Try to keep assay plate in a sealed aluminium foil bag and avoid dampness.
Restrictions
For Research Use only