ZipTip Protocol for MALDI-TOF
Solutions Required
0.5% TFA in HPLC water
- 1 mL: 5 μL of TFA in 995 μL of HPLC water
50:50 acetonitrile:water
100% acetonitrile (ACN)
Matrix CHCA (Final concentration: 6.7 mg/mL; 67% ACN; 33% water; 0.2% TFA). To a 5 mg CHCA tube add:
- 500 μL of ACN
- 250 μL of 0.5% TFA in water
This protocol is used to remove salts from the sample. The ZipTip can trap up to 2 μg of peptides/proteins.
Procedure
- Concentrate/dry the samples in the speedvac to a volume of 10 μL or less.
- If sample volume gets too low (less than 5μL) after speedvac concentration, add 5-10μL of 0.5% TFA in HPLC water.
- Make 1.5 μL aliquots of matrix CHCA into desired number of tubes (depends on number of samples).
- Vortex and centrifuge sample tubes.
- Place ZipTip on 10μL or 20μL Pipetmen.
- Wash ZipTip 2x with Acetonitrile solution (ACN).
- Wash ZipTip 1x with 50:50 ACN:H20
- Wash ZipTip 2x with 0.5% TFA solution.
- Run sample through ziptip 7-10x SLOWLY.
- Wash sample 1x with 0.5% TFA H2O SLOWLY.
- Load elution buffer (matrix CHCA aliquot of step 3) onto ZipTip.
- Elute sample onto target (spot on plate).
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