Kaushik Shandilya
Prepare the DTT solution: Prepare a 0.1 M DTT solution in phosphate-buffered saline (PBS). The solution should be freshly prepared and kept in the dark. Collect the airborne bioaerosols: Collect the bioaerosols using an appropriate sampling device such as a high-volume air sampler or an impinger. Ensure that the sampling device is properly cleaned and sterilized before use. Extract the bioaerosols: Extract the bioaerosols from the collection substrate using an appropriate extraction method such as sonication or vortexing. Prepare the reaction mixture: Prepare a reaction mixture by adding the extracted bioaerosols to the DTT solution. The ratio of bioaerosols to DTT should be optimized for each sample to ensure the best assay performance. Incubate the reaction mixture: Incubate the reaction mixture at 37°C for 1 hour. The reaction should be carried out in a dark environment to prevent any interference from light. Stop the reaction: Stop the reaction by adding an acidic stop solution (such as trichloroacetic acid) to the reaction mixture. The stop solution should be added in a 1:1 ratio to the reaction mixture. Analyze the samples: Analyze the samples using a spectrophotometer to measure the absorbance at 412 nm. The absorbance value reflects the amount of DTT consumed by the bioaerosols, which is an indicator of their OP.
Dear friend Akshay Kale
The DTT (dithiothreitol) assay is a commonly used method to assess the oxidative potential (OP) of airborne bioaerosols. Here are the general steps involved in conducting the DTT assay for OP:
To ensure the accuracy of the assay, it is important to include appropriate controls such as a blank control (containing only the reaction mixture without any bioaerosols) and a positive control (containing a known oxidative agent such as hydrogen peroxide).
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