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TMB (3,3',5,5' Tetramethylbenzidine)

TMB (3,3',5,5' Tetramethylbenzidine)

$258.00

Key features and details

3,3′,5,5′-Tetramethylbenzidine (TMB) is a noncarcinogenic substitute (Ames test negative) for benzidine suitable as peroxidase substrate for use in ELISA procedures. The substrate produces a soluble end product that is pale blue in color and can be read spectrophotometrically at 370 or 620-650 nm. The TMB reaction may be stopped with 2 M H2SO4 (resulting in a yellow color), and read at 450 nm. A sensitive and specific reagent for the detection of blood, assay of hemoglobin, assay of peroxidases.

SKU: 0898050-CF

CAS Number:
54827-17-7
Molecular Weight:
240,35
Product Description

3,3′,5,5′-Tetramethylbenzidine (TMB) is a noncarcinogenic substitute (Ames test negative) for benzidine suitable as peroxidase substrate for use in ELISA procedures. The substrate produces a soluble end product that is pale blue in color and can be read spectrophotometrically at 370 or 620-650 nm. The TMB reaction may be stopped with 2 M H2SO4 (resulting in a yellow color), and read at 450 nm. A sensitive and specific reagent for the detection of blood, assay of hemoglobin, assay of peroxidases.

Application Notes

TMB is a substrate for horseradish peroxidase. Develops a soluble blue reaction product that may be read at 370 or 655 nm. For endpoint assays the reaction may be stopped with acid, forming a yellow reaction product that may be read at 450 nm. Recommended for ELISA (microwell) procedures, not recommended for membrane applications.

Protocol to make TMB stock solution and working solution:

TMB Stock Solution: 50 mg/mL in DMSO

100 mg TMB (SKU 08980502 or 08980501), dissolve it in 2 mL DMSO (You can scale up).

Store it at -20 °C. The solution should be active for one year. Any oxidation would be revealed

as a color change in the working solution.

TMB working solution: 0.05%

50 μL of 50 mg/mL TMB stock solution in DMSO

5 mL of phosphate-citrate buffer, pH 5.0

Phosphate-citrate buffer, pH 5.0

Prepare 800 mL of distilled water in a suitable container

Add 14.62 g of Disodium Hydrogen Phosphate to the solution

Add 10.19 g of Citric Acid to the solution

Adjust solution to final desired pH using phosphate acid or NaOH

Add distilled water until volume is 1 L

Usage Statement

Unless specified otherwise, MP Biomedical's products are for research or further manufacturing use only, not for direct human use. For more information, please contact our customer service department.

Key Applications

ELISA

Specifications
SKU 0898050-CF
Application Notes TMB is a substrate for horseradish peroxidase. Develops a soluble blue reaction product that may be read at 370 or 655 nm. For endpoint assays the reaction may be stopped with acid, forming a yellow reaction product that may be read at 450 nm. Recommended for ELISA (microwell) procedures, not recommended for membrane applications.

Protocol to make TMB stock solution and working solution:

TMB Stock Solution: 50 mg/mL in DMSO

100 mg TMB (SKU 08980502 or 08980501), dissolve it in 2 mL DMSO (You can scale up).

Store it at -20 °C. The solution should be active for one year. Any oxidation would be revealed

as a color change in the working solution.

TMB working solution: 0.05%

50 μL of 50 mg/mL TMB stock solution in DMSO

5 mL of phosphate-citrate buffer, pH 5.0

Phosphate-citrate buffer, pH 5.0

Prepare 800 mL of distilled water in a suitable container

Add 14.62 g of Disodium Hydrogen Phosphate to the solution

Add 10.19 g of Citric Acid to the solution

Adjust solution to final desired pH using phosphate acid or NaOH

Add distilled water until volume is 1 L

Base Catalog Number 980501,980502
CAS # 54827-17-7
Formulation Details Powder
Molecular Weight 240,35
Usage Statement Unless specified otherwise, MP Biomedical's products are for research or further manufacturing use only, not for direct human use. For more information, please contact our customer service department.
Citations