단백질 기반 연구

제품문의 관련제품보기 관련내용보기

Sample Processing on the PhyNexus MEA2 Personal Purification System

Part I. Trypsin PhyTip digestion.

1. 10 μL samples were placed in dark 96-well plate in Position 4. Plate was covered with foil sticker.
2. Denaturing: Pick up 20 μL 8M Guanidine-HCl and add into sample wells.
3. Reduction: Pick up 5 μL DTT and add into sample wells, react for 30 min at RT.
4. Alkylation: Pick up 5 μL IAA and add into sample wells, react for 20 min at RT.
5. Dilution: Pick up 60 μL 50 mM Ammonium Bicarbonate from Channel C at position 8 and add into sample wells. Final sample volume is 100 μL.
6. Trypsin tip equilibration: Pick up trypsin tips at position 1, equilibrate tips in Channel D at position 8.
7. Trypsin digestion: Move trypsin tips into sample well and perform on-column digestion at RT.
8. pH adjustment: Pick up 7μL 5% TFA to acidify samples for desalting.

Part II. C18 PhyTip desalting.

1. Buffer preparation: Pick up 100 μL 0.1% TFA and dispense into 96-well plate at position 5. Then pick up 100 μL elution buffer and dispense into 96-well plate at position 7.
2. C18 tip activation: Pick up C18 tips at position 2, activate tips in Channel F at position 8.
3. C18 tip equilibration:Move C18 tips into Channel G at position 8, equilibrate tips in 0.1% TFA.
4. Peptide binding: move C18 tips into sample wells, perform peptide binding procedure.
5. C18 tip washing: move C18 tips into wash plate at position 5, wash tips in 0.1% TFA.
6. Peptide elution: move C18 tips into elution plate at position 7, elute peptides in 60% ACN, 0.1% TFA.