How to Perform an Absorption Based Assay

Step-by-Step Guide

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   Step 1: Read the assay instruction sheet before starting the experiment.

     Gather all materials and equipment needed to perform the experiment.
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   Step 2: Plan out how much of each material will be needed to obtain the desired concentrations and which wells of the microplate to put it in.

   Follow the instruction sheet in order to create the right mixtures of chemicals. , Based on the instruction sheet of the specific experiment. ,,, In order to understand technical and biological variations within the experimental system, use replicate samples.
   
   Have a row or column of just the chemical solvent to serve as a control solution. ,,, It will be one of the following: end point, kinetic, spectrum , Adjust the range of absorbance readings as needed for the specific assay , It is important to set the proper microplate setting to match the microplate being used. ,, Determine the intensity and duration of shaking. , Determine which wells to read. ,
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   Step 3: Add in the necessary reagents (enzymes

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   Step 4: buffer solution

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   Step 5: Use a microplate that is compatible with the microplate reader.

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   Step 6: Dilute stock solutions prior to pipetting.

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   Step 7: Using a micropipette

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   Step 8: deposit the desired amount of reagents into the wells of a microplate.

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   Step 9: Create a protocol to fit your assay by adjusting the microplate reader’s settings according to the manufacturer’s operating manual.

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    Step 10: Read the instruction plate before adjusting the microplate reader settings

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    Step 11: Adjust the reading type.

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    Step 12: Adjust the wavelengths Adjust the range of wavelengths for excitation and emission as needed for the specific assay.

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    Step 13: Specify the type of microplate There are multiple types of microplates (48-well

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    Step 14: 96-well

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    Step 15: etc.).

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    Step 16: Adjust temperature control

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    Step 17: if needed Temperature level – Enter the temperature required for the experiment Preheating Option – If needed

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    Step 18: click the pre-heating option so that the microplate reader will preheat to the correct temperature Add in the incubation time.

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    Step 19: Adjust shaking settings

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    Step 20: if needed.

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    Step 21: Set up the layout of the plate.

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    Step 22: Set the microplate into the microplate reader and start analysis.

Detailed Guide

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