Portelite™ Fluorimetric Lithium Ion Quantification Kit

Thaw all the kit components at room temperature before starting the experiment.

1. Prepare the Lithiumighty™ 520 working solution.

2. Add 100 µL of the Lithiumighy™ 520 working solution to each 0.2 mL PCR tube.

3. Add 100 µL of the Lithium Standards or test samples into each tube.

4. Incubate at room temperature for 5-10 minutes.

5. Monitor the fluorescence intensity with a CytoCite™ fluorometer or Qubit™ fluorometer.

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

1. To prepare a Lithiumighty™ 520 stock solution, add 100 μL of DMSO to the vial containing Lithiumighty™ 520 (Component A).

   Note: Prepare a single aliquot of unused Lithiumighty™ 520 stock solution and store it at ≤ -20 ºC, protected from light. Avoid freeze/thaw cycles.

1. Prepare the Lithiumighty™ 520 working solution by adding 100 μL of Lithiumighty™ 520 (Component A) to 5 mL of Assay Buffer (Component B), and protect the working solution from light by covering it with foil or placing it in a dark location.

   Note: For optimal results, use this solution within a few hours after preparing it.

   Note: 5 mL of working solution is enough for 100 tests.

The acceptable sample volume can range from 1 to 20 μL, depending on the estimated concentration of the sample. The following protocol is based on a sample volume of 10 μL.

1. Add 100 µL of Lithiumighty™ 520 working solution to each Cytocite™ sample tube (Cat No. CCT100) or an equivalent 0.2 mL PCR tube.

2. Add 100 μL of Lithium Standards or test samples to each tube. Mix each tube by vortexing for 2-3 seconds.

3. Incubate all the tubes at room temperature for 5-10 minutes.

4. Insert the samples into either the CytoCite™ or Qubit™ devices. Use the green fluorescence channel to measure the fluorescence intensity. Be sure to follow the specific procedures for the CytoCite™ Fluorometer. For detailed instructions, refer to the link below:

   https://devices.aatbio.com/documentation/user-manual-for-cytocite-fluorometer

For Portelite™ assays, you can create a calibration curve using the Lithium Standards. Below is a simple protocol for generating a customized Lithium standard curve.

1. Perform a 1:2 serial dilution. First, add Lithium Standard (Component C) into the Assay Buffer (Component B). Then, create the following Lithium standard dilutions: 300 mM, 150 mM, 75 mM, 37.5 mM, 18.75 mM, 9.375 mM, and 4.6875 mM.

   Note: Final, in well concentration of the sample, will be 150, 75, 37.5, 18.75, 9.37, 4.68, 2.34 mM.

2. Add 100 µL of the Lithiumighty™ 520 working solution to each tube.

3. Add 100 µL of either standards or samples into a 0.2 mL PCR tube.

4. Incubate the reaction at room temperature for 2 minutes.

5. Insert the samples into the CytoCite™ device and use the green fluorescence channel to monitor the fluorescence intensity.