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.