Thermophoresis, Microscale

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Monolith NT.115 (B413)

CAPTAIN: Pfeffer Lab - B413

Rules and Guidelines

1. Register on the calendar to use the machine.

2. If you are the last person of the day, shut the computer down and turn off the machine (switch on back left).

3. Clean out capillaries when done with your run.

4. Turn off temp. control when done with your run if the next person does not require it.

5. Clean up the bench when done.

Before use:

1. New users MUST be trained by equipment captain.

2. For detailed instructions, please read the manual below.

3. Please register for time on the indicated Google Calendar with your name and room number. If you need temperature control add Temp and the Temp setting you need. i.e. “Tory Johnson B413 Temp 22C”. An MST run at a single power level will take ~25 min.

Instructions for Use:

1. Turn on machine (power switch back left) and computer. Machine does not need to warm up.

2. Open NT Control application on desktop to start program.

3. Start a New Project and save it to C:\users\nanotemper\documents\[Your Lab]\[Your Name]\

4. The up button on the touchscreen of the machine opens the machine.

5. Load your samples in capillary tubes [do not touch the center of the tubes].

6. Load sample chamber into machine and close the machine (down button on touchscreen).

7. Set the LED color and power level.

8. Set the dilutions used across your series.

9. Cap scan button determines the position of the capillary tubes as well as signal of label. Ideally, signal should be between 200-2000.

10. Set MST power and time (0-50% power, start with 30s; 50-100% start with 20s).

11. Once done experiment can be analyzed with Affinity analysis app on desktop.

Equipment Description:

The Monolith NT.115 is equipped with Nano-BLUE/RED fluorescent channels (compatible with AF488,AF647). The Monolith NT.115 allows measuring every interaction type - ranging from the formation of huge protein

complexes to the binding of single metal ions. As there are no prerequisites with respect to the buffer composition, even such complex liquids as serum or lysates can be used for interaction studies (see for more information).



START GUIDE: File:StartingGuide NT115-2.pdf

TRAINING GUIDE: File:MST Training Stanford-2.pdf


File:FAQ Buffer composition V06-2.pdf

File:FAQ Different Dyes V06-2.pdf

File:FAQ Fluorescence Changes V11-2.pdf

File:FAQ NT.115 Filtersets V03-2.pdf

File:FAQ Stoichiometry V03-2.pdf

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