Stop Solution for TMB Substrate (650nm, non-corrosiveness)

Stop Solution for TMB Substrate (650nm, non-corrosiveness) is a safe, non-corrosive, ready-to-use solution mainly used to terminate TMB chomogenic reactions in ELISA and other experiments. After the addition of this product, the TMB reactions is terminated, but with no colour change, and the absorbance can be measured at 620-650nm.This product is very safe, does not contain sulfuric acid or other medium to strong acids, which effectively avoids the potential harm by the strong corrosiveness of sulfuric acid to the human body.TMB, namely 3, 3',5 ,5'-Tetramethylbenzidine, is a common substrate of horseradish peroxidase (HRP). Under the catalysis of HRP or other appropriate peroxidases, TMB produces a soluble blue product with an absorbance at 370nm or 620-650nm. The TMB chromogenic reactions need to be terminated in time. Otherwise, the experimental results may be affected because the reaction between HRP and TMB can continue and reach a plateau after a certain period of time. Stop Solution for TMB Substrate is divided into two types according to the absorbance, one at 450nm and the other at 620-650nm. The color of TMB reactions does not change after the addition of this product, and the absorbance which is stable for 1 hour can be measured at 650nm. Please refer to Figure 1 for the assay results after the addition of this product.Figure 1. Reactions of HRP at different concentrations with the TMB Substrate Solution for ELISA were terminated by the addition of Stop Solution for TMB Substrate (650nm, non-corrosiveness) . The absorbance was measured at 370nm before the addition of this product and at 650nm after the addition of this product, respectively. Left panel, A370 and A650 of 5-minute reactions at room temperature; Right panel, Color of reactions before (upper) or after (lower) the addition of this product. This figure is for reference only, which may vary due to different experimental conditions.The comparison of Aladdin's Stop Solution for TMB Substrate (450nm, Sulfuric acid free) and Stop Solution for TMB Substrate (650nm, non-corrosive) is shown in the following table.Cat. No.P0215 P0217 Product Name Stop Solution for TMB Substrate (450nm, sulfuric acid free)Stop Solution for TMB Substrate (650nm, non-corrosive)Final Reaction Color YellowBlueDetection wavelength 450nm 620-650nm Absorbance SignalApproximately doubledNo significant changeSafetyCorrosiveNon-corrosivePrecautions：This product is suitable for the termination of TMB chomogenic reactions in solution, such as ELISA, but not suitable for histochemical or blotting assays.After adding TMB Substrate Solution or Stop Solution for TMB Substrate, the occurrence of precipitation indicates that the sample contains too high level of target protein and should be diluted appropriately before the assay.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1. For ELISA: a. (Refer to the instructions of a ELISA kit) after incubation with HRP-labeled antibody, wash 3-5 times with an appropriate wash buffer for 3-5 minutes each. b. Remove the wash buffer and add 100µl of TMB Substrate Solution. c. Incubate at room temperature in the dark for 3-30 minutes or longer until the expected color intensity is achieved. d. Add 100µl of Stop Solution for TMB Substrate (650nm, non-corrosiveness), then measure the absorbance at 650nm.2. For other assays in 96-well plates (e.g., detection of endogenous peroxidase in cell or tissue samples):a. Add 10-20µl of sample directly to each well of 96-well plates. b. Add 100µl of TMB Substrate Solution. c. Incubate at room temperature in the dark for 3-30 minutes or longer until the expected color intensity is achieved.d. Add 100µl of Stop Solution for TMB Substrate (650nm, non-corrosiveness), then measure the absorbance at 650nm.

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