Introduction
TMB solutions are chromogenic reagents for peroxidase, used for ELISA techniques. They are composed of 3,3’,5,5’-tetramethylbenzidine (TMB), hydrogene peroxide (H2O2), and proprietary catalyzing and stabilizing agents. It produces a blue product upon interaction with HRP or HRP conjugates without the addition of hydrogen peroxide. The soluble blue product can be quantitated at 650 nm. Sensitivity is greater than classic substrates like OPD and ABTS, with very low background.
Storage
2-8°C avoid light for three years. Do not freeze (Return TMB Solution to room temperature before use).
Seize: 10mL, 50 mL, 100mL,1000mL,5000mL.
ELISA Applications
1. Wash the microplate following the incubation of HRP-labeled solution.
2. Add 100 µL TMB Solution into each well. Incubate the plate at 37°C for 15–30 min or until the
desired color develops.
3. Measure the absorbance signal at 650 nm with an ELISA microplate reader.
Note: If desired, the reaction can be stopped by adding an equal volume of 2M sulfuric acid to
each well. Stopped reaction should be read at 450 nm.
NOTE:
1. TMB substrates are optimized for direct and indirect ELISA techniques. They are not suitable for
2. Immunohistochemistry or Western Blotting.
3. Recommendations for use: TMB is sensitive to certain conditions of storage and operating.
4. Exposure to light : TMB substrate is light sensitive. It should be stored in amber vials of HDPE
type.
5. Material : the substrate is very sensitive to metallic ions. Only high quality plastic or glass should
be used.
6. Avoid the use of caps with rubber joins : this should impair the results.
7. Don’t pipet the TMB directly from the bottles : fill a suitable container first with the necessary
solution volume to avoid contamination.
8. Don’t leave the bottle open too long , and keep it a dark at 2-84°C. Avoid important or
frequent variations of temp.
Ecalbio Co.,Ltd specialize in developing reagents of immunodiagnostic technology and biological systems with higher accuracy andreliability for results of the highest quality.
Ecalbio builds up professional platform to develop more than 10,000 products, including antibodies, antigens, biochemicals, ELISA kits, CLIA kits, and laboratory equipments,etc.
Ecalbio laboratories manufacture Antibodies, Proteins, Food safety ELISA kits, Food safety Rapid Tests, Animal Disease ELISA Kits, Animal Disease Rapid Test Kits, Laboratory equipments with incubators and diagnostic system.
Ecalbio has an enthusiastic and creative R&D team, consisting with a group of experienced technicians in lateral flow immunoassay field, and set up long relationship with first-class institutes,universities,academies, to provide excellent service for human healthy and earth.
Ecalbio are committed to saving lives by ensuring individuals worldwide have access to timely and correct diagnosis. Our products are used in over 100 countries worldwide by:
Government Laboratories
Food industry Laboratories
Institutes
Veterinary Clinics
Pharmaceutical Industry.
Ecalbio welcomes potential partners and distributors to explorebusiness relationships with all customer in the worldwide, focused on scie-tech, integrity, quality and service for the customers.
ECALBIO CO.,LTD
HZAU, Luoshi road, Wuhan 430060, China
Tel:86-27-87745856
Email:info@ecalbio.com
Website: www.ecalbio.com