产品名称：O-PHENYLENEDIAMINE, DIHCL, 10-MG TABLETS
Application Notes: 0
Note: The tablets are inspected visually and are examined spectrophotometrically at 492 nm for the background level in 0.1 M sodium phosphate/citric acid, pH 5.0, containing 6 mM H2O2. Tablets dissolve in the substrate buffer within 5 min (with shaking).
Suggested Procedure for Use of OPD tablets for HRP-based ELISAs:
REAGENTS REQUIRED BUT NOT PROVIDED:
1. Substrate buffer: 50 mM citric acid/sodium phosphate buffer, pH 5.0.
2. Hydrogen peroxide (H2O2) 30%.
3. Stop solution: 0.1 M H2SO4.
1. Complete all required incubations with antibodies and HRP-labeled probes.
2. Wash plate wells at least 4 times with phosphate-buffered saline (PBS) or Tris-buffered saline, containing 0.1% TWEEN®-20 detergent.
3. After the final wash, shake and blot all residual buffer from the wells.
4. Within 30 min of completion of the above steps, prepare OPD Substrate Solution (the following recipe provides sufficient OPD Substrate Solution for one 96-well plate (100 μl per well):
a. Remove the OPD tablet container from the refrigerator about 15 min before use.
b. Dissolve 1 tablet (10 mg total) in 12.5 ml of Substrate Buffer at room temperature with stirring. Note: DO NOT use metal forceps during handling of the tablets, as metals catalyze the oxidation process. Protect the solution from direct sunlight.
c. Add 12.5 μl of 30% H2O2, mix, and use within 30 min. [Final H2O2 concentration = 0.03% (v/v)]
5. Add 100 μl of OPD Substrate Solution to each well, and incubate 5-30 min.
Note: Optimal incubation times may vary depending on the amount of HRP present. If color develops too quickly, dilution of the probe, antibody, or HRP-labeled reagent may be required.
6. Stop reaction by adding 100 μl of 0.1 M H2SO4.
7. Read the absorbance at 492 nm.
Note: Variations in incubation times and reagent concentrations may require further standardization by the user.
Presentation: Off-white tablets; White tablets (10 mg each), containing no preservative.
Storage Conditions: Close the lid of the container tightly for storage, and do not remove the enclosed drying capsule. Protect from light.