Maldi-TOF / TOF
General Instructions for MALDI TOF/TOF Users:
Mass spectrometry is highly sensitive to various contaminants, such as PEG (poly ethylene glycol), keratin and various salts. In order to reduce PEG and keratin in the sample it is recommended to use powder free nitryl gloves. Please supply the sample in a low-salt buffer (less than 10 mM).
- Use fresh solutions. All solvents should be pure as possible. HPLC grade is recommended.
- Avoid volatile solvents! Several of these solvents are common in protein chemistry: glycerol, polyethylene glycol,b-mercaptoethanol,triton-X-100,DMSO,DMF,etc.
- Scalpel, pipette tips and tubes should be clean. Use original Eppendorf tubes. Since the solvents can leach the plastics and polymer contamination can occur.
- Sequencing grade trypsin is recommended
It is recommended to use the Coomassie / silver staining Standard protocol.
Levels of buffers and detergents which exceed the following limits will probably cause noticeable degradation of the spectrum
| Phosphate Buffer | >50 mM |
| Ammonium Bicarbonate | > 30 mM |
| Tris Buffer | >100 mM |
| Guanidine | >1M |
| Detergent(e.g. Trition-X) | 0.1% |
| SDS | 0.01% |
| Alkali metal salts | >1M |
| Glycerol | >1% |
| Sodium azide | >1 mM |
Contact Details:
Email: maldi.bios@nulliitb.ac.in
Contact Phone # – 022 25764757