523120 | o-Phenylenediamine, Dihydrochloride, 2-mg Tablets

523120
  
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      Overview

      Replacement Information

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      Description
      Overview

      This product has been discontinued.





      Contains 50 tablets. Chromogenic substrate for horseradish peroxidase in ELISA procedures.
      Catalogue Number523120
      Brand Family Calbiochem®
      SynonymsOPD, o-PD, HCl
      References
      Product Information
      CAS number615-28-1
      FormOff-white tablets
      Hill FormulaC₆H₈N₂ . 2HCl
      Chemical formulaC₆H₈N₂ . 2HCl
      Applications
      Application CommentsThe tablets are inspected visually and are examined spectrophotometrically at 492 nm for background levels 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

      PROTOCOL:
      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®-detergent 20.
      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 5 tablets (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.
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      RTECSST0175000
      Safety Information
      R PhraseR: 23/24/25-43-50/53

      Toxic by inhalation, in contact with skin and if swallowed.
      May cause sensitization by skin contact.
      Very toxic to aquatic organisms; may cause long-term adverse effects in the aquatic environment.
      S PhraseS: 28-36/37-45-60-61


      Wear suitable protective clothing and gloves.
      In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible).
      This material and its container must be disposed of as hazardous waste.
      Avoid release to the environment. Refer to special instructions/safety data sheet.
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Ambient Temperature Only
      Toxicity Toxic & Carcinogenic / Teratogenic
      Hazardous Materials Attention: Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
      Storage +2°C to +8°C
      Protect from Light Protect from light
      Do not freeze Ok to freeze
      Special InstructionsClose the lid of the container tightly for storage, and do not remove the enclosed drying capsule.
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      523120

      Brochure

      Title
      Enzyme-Linked Detection Systems Technical Bulletin
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision10-August-2007 RFH
      SynonymsOPD, o-PD, HCl
      Descriptiono-Phenylenediamine (OPD) dihydrochloride supplied as 2 mg tablets.
      BackgroundOPD (ortho-phenylenediamine; o-PD) is a sensitive, chromogenic substrate for horseradish peroxidase (HRP)-based enzyme-linked immunosorbent assays (ELISA). In the presence of HRP and hydrogen peroxide (H2O2), OPD is oxidized to a water-soluble, yellow-orange product that is measured spectrophotometrically at 492 nm, allowing a quantitative analysis of HRP-based systems.
      FormOff-white tablets
      CAS number615-28-1
      RTECSST0175000
      Chemical formulaC₆H₈N₂ . 2HCl
      CommentsThe tablets are inspected visually and are examined spectrophotometrically at 492 nm for background levels 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

      PROTOCOL:
      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®-detergent 20.
      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 5 tablets (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.
      Storage +2°C to +8°C
      Protect from light
      Do Not Freeze Ok to freeze
      Special InstructionsClose the lid of the container tightly for storage, and do not remove the enclosed drying capsule.
      Toxicity Toxic & Carcinogenic / Teratogenic
      Merck USA index14, 7284