One Component TMB Color Development Solution(ELISA&HRP)

ELISA, also known as enzyme-linked immunoassay, is a widely utilized method for both qualitative and quantitative analysis of antigens, haptens, and antibodies. In this technique, horseradish peroxidase (HRP) and its conjugates are commonly employed to catalyze the chromogenic reaction of substrates such as 3,3'5,5'-tetramethylbenzidine (TMB), enabling accurate analysis. TMB is highly sensitive and free from carcinogenic properties, making it the preferred choice for various experiments, including ELISA, immunoblot hybridization, immunohistochemistry, and even chlorine detection.
However, traditional TMB colorimetric reagents consist of multiple components that need to be prepared individually, leading to the potential for precipitation and unstable test results. To address these issues, a novel TMB color technology has been developed. This innovative approach involves combining all the necessary reagents in a single solution, simplifying the process and ensuring more reliable and consistent test outcomes. Specifically designed for ELISA experiments, this product operates on the principle that HRP or another appropriate peroxidase can facilitate the formation of soluble blue products, which can be measured at 370nm. The chromogen reaction is halted by the addition of an acidic solution (e.g., 0.5-2m H2SO4, 0.5M HCl, 1M H3PO4), causing the blue product to transition into a yellow form. The absorbance of the yellow solution can then be measured at 450nm.
Thanks to its superior color sensitivity and wide linear concentration range, this product proves exceptionally suitable for quantitative ELISA analysis. Typically, for ELISA detection, each sample requires 0.1ml of the chromogenic solution. Therefore, a 100ml quantity of this product allows for the analysis of approximately 1000 samples, while 500ml enables the examination of around 5000 samples. This streamlined and efficient solution simplifies the ELISA process, ensuring accurate and consistent results every time.

HOW TO USE (ELISA)
(2) Before using the single-component TMB chromographic solution, it is necessary to take out an appropriate amount and allow it to return to room temperature. This will ensure its effectiveness during use.
Once the ELISA detection kit has been incubated with the HRP-labeled antibody for a specific amount of time, it is necessary to wash it thoroughly with a suitable detergent. This should be done at least three to five times, with each wash being performed for a duration of three to five minutes. This step becomes crucial in ensuring that any unbound components are removed, and only the desired results can be obtained. Thus, washing helps in attaining the accuracy and specificity of the test.
(3) After washing, remove the washing solution and add 100μl one-component TMB color developing solution.
(4) Incubate at room temperature and avoid light for 5-30min or longer (up to 24h) until the color reaches the expected depth.[Note] : It is recommended to choose the best reagent dosage, color developing time and reaction temperature according to the experimental requirements.
(5) Determination of absorbance directly at 370nm.Alternatively, add 100μl 0.5-2m H2SO4 to terminate the reaction and determine the absorbance at 450nm.

Store at 2-8° C avoid light for 24 months

1. This product contains TMB and H2O2, which is irritant to human body. Please pay attention to protection.
2. If the product is found to be cloudy or blue in color, please stop using it.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.

TMB One Solution (HRP-based ELISA)