Serum Total Cholesterol Assay: Difference between revisions
added clarifying notes |
added directions for plate reader |
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* In the middle of the time add the standards (2, 4, 6, 8 uL of 10x and 1, 1.5, 2 uL of the undiluted cholesterol standard) | * In the middle of the time add the standards (2, 4, 6, 8 uL of 10x and 1, 1.5, 2 uL of the undiluted cholesterol standard) | ||
* The assay takes 15 minutes to develop at room temperature, and is stable for 30 minutes. That means if the timer hits 30 minutes you wont be able to finish in time, so stop and start the plate. | * The assay takes 15 minutes to develop at room temperature, and is stable for 30 minutes. That means if the timer hits 30 minutes you wont be able to finish in time, so stop and start the plate. | ||
* Once all samples are added, incubate plate at room temperature for | * Once all samples are added, incubate plate at room temperature for 15 minutes | ||
* Measure absorbance at 500 nm | |||
=== Plate Reader === | |||
*Turn on computer, screen, and plate reader (switch on back right) | |||
*Log in | |||
*Open SoftMaxPro 5.3 | |||
*Measure absorbance at 500 nm (Under settings, set Lm1 to 490. Our machine does not have a filter for 500 nm) | |||
*Click Read | |||
*File —> Print | |||
*Save —> FileName YYYY-MM-DD-Chol | |||
*Save in Bridges Folder | |||
*Export —> samename.txt | |||
*Get txt file via USB drive and print | |||
== Calculations == | == Calculations == | ||