ZipTip Protocol for LC-MS/MS
Solutions Required
0.5% TFA in HPLC water
- 5 μL of TFA in 995 μL of water
50:50 acetonitrile:water
100% Acetonitrile (ACN)
Elution Buffer (67% ACN; 32.8% Water; 0.2% TFA)
- 670 μL of 100% acetonitrile, 328 μL of water, and 2 μL of TFA
Solvent A: 0.1% formic acid in water
This protocol is used to remove salts from the sample. The ZipTip can trap up to 2 μg of peptides/proteins.
Procedure
- Dry down samples in the SpeedVac.
- Resuspend the samples in 20 μL of solvent A.
- Aliquot 20 μL of elution buffer into desired number of tubes.
- Using the ZipTip, wash 2X with acetonitrile solution (ACN).
- Wash ZipTip 1X with 50:50 ACN:H2O
- Wash ZipTip 2X with 0.5% TFA solution.
- Run sample through ZipTip 10X SLOWLY.
- Wash sample 1x with 0.5% TFA in Water SLOWLY.
- Wash ZipTip 10X with the 20 μL aliquot of elution buffer.
- Dry down the sample using the SpeedVac. Once dry, resuspend the sample with 7 μL of A.
- Repeat as necessary with remaining samples.
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