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			96-Well Plate

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	48-602MAG	Buffer Detection Kit for Magnetic Beads	1 Kit

523121 Sigma-Aldricho-Phenylenediamine, Dihydrochloride, 10-mg Tablets

o-Phenylenediamine, Dihydrochloride, 10-mg Tablets MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

SDS
CoA
Data Sheet

Synonyms: OPD, o-PD, HCl

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Overview

Replacement Information

Pricing & Availability

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Description
Overview	This product has been discontinued. We are offering [insert product name] (Cat. No. P8287) as a possible alternative. Please read the alternative product documentation carefully and contact technical service if you need additional information. Contains 50 tablets. Chromogenic substrate for horseradish peroxidase in ELISA procedures.
Catalogue Number	523121
Brand Family	Calbiochem®
Synonyms	OPD, o-PD, HCl

References

Product Information
CAS number	615-28-1
Form	Off-white tablets
Formulation	White tablets (10 mg each).
Hill Formula	C₆H₈N₂ . 2HCl
Chemical formula	C₆H₈N₂ . 2HCl
Preservative	None

Applications
Application Comments	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. 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^®-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.

Applications

Application Comments

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.

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^®-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.

Biological Information

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS
RTECS	ST0175000

Safety Information
R Phrase	R: 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 Phrase	S: 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	Blue Ice 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	-20°C
Protect from Light	Protect from light
Do not freeze	Ok to freeze
Special Instructions	Close the lid of the container tightly for storage, and do not remove the enclosed drying capsule.

Packaging Information

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
523121-1EACN	07790788051037

Documentation

o-Phenylenediamine, Dihydrochloride, 10-mg Tablets SDS

Title
Safety Data Sheet (SDS)

o-Phenylenediamine, Dihydrochloride, 10-mg Tablets Certificates of Analysis

Title	Lot Number
523121	Enter Lot Number

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.

Revision	10-August-2007 RFH
Synonyms	OPD, o-PD, HCl
Description	OPD (ortho-phenylenediamine) tablets for preparation of soluble substrate.
Background	OPD (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.
Form	Off-white tablets
Formulation	White tablets (10 mg each).
CAS number	615-28-1
RTECS	ST0175000
Chemical formula	C₆H₈N₂ . 2HCl
Preservative	None
Comments	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. 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^®-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.
Storage	Protect from light -20°C
Do Not Freeze	Ok to freeze
Special Instructions	Close the lid of the container tightly for storage, and do not remove the enclosed drying capsule.
Toxicity	Toxic & Carcinogenic / Teratogenic
Merck USA index	14, 7284

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