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One-Component TMB Substrate (for ELISA)
The main component of the TMB substrate solution is 3,3',5,5'-tetramethylbenzidine (TMB), which is horseradish peroxidase (HRP). The substrate, under the action of horseradish peroxidase, reacts with an oxidizing agent to form a blue product. The intensity of the color is directly proportional to the activity of HRP and can be used to detect HRP-based detection.
The ELISA is used for the labeling of horseradish peroxidase; the single component, the color developing solution is ultra-stable, the solution is also quite stable after termination, and the use is more convenient and the sensitivity is high.
The product packaging specifications are 100ml, 500ml, 1000ml
Transportation, storage and expiration date
Store at room temperature (do not exceed 35 ° C), store at 2-8 ° C, use within the validity period. Under normal storage conditions, the validity period is 2 and a half years from the date of production.
This product is suitable for ELISA experiments with HRP label. TMB and hydrogen peroxide contained in this product. Under the action of HRP, TMB is oxidized to form water-soluble blue product with absorption peak at 370nm and 650nm. When performing kinetic experiments, the absorbance at 650 nm can be directly detected. End point detection can be performed by adding a stop solution. The stop solution was a 0.5 M sulfuric acid solution or a 1 M hydrochloric acid solution. After adding the stop solution, it turns yellow, the detection wavelength is 450 nm, and the absorbance value can be increased several times compared with the non-acidification.
1. This product is sensitive to various oxidants. It should be avoided during use. Do not insert the tip directly into the bottle to take the liquid. The required amount of liquid should be poured out. Do not return the remaining solution to the original bottle. Pollution.
2. Avoid direct sunlight during operation. It is recommended to use plastic containers for the containers to avoid the use of metal containers and containers contaminated with bacteria and redox agents.
3. Excessive concentrations of HRP can cause TMB to appear purple, or precipitation after termination, which should reduce the amount of antibody and label or shorten the incubation time.
Method of operation
1. Add 100 μl of TMB solution to each well
2. Incubate for 5 to 30 minutes at room temperature, adjust the reaction time according to the needs of the reaction system, and make the absorbance value after termination to the appropriate range.
3. Add 50 μl stop solution (0.5 M H2SO4 or 1 M HCl).
Detection within 30 minutes after termination, detection on a microplate reader, detection wavelength is 450 nm.