Instructions for Use - Glyco Kit MB-LAC AIA - Bruker

More documents

Recommendations

Info

General remarks for sample handling and MALDI measurements MALDI-TOF mass spectrometry is a very sensitive analysis method requiring special operation procedures. Therefore, we recommend considering the following aspects before starting an analysis: The quality of the starting material will have an enormous impact on the outcome of the experiments. Especially, collection and storage conditions have to be controlled carefully. When working with clinical samples, like serum or urine, please refer to the recent publications of Baumann et al. 8 and Fiedler et al. 9 for standardized sample handling procedures. To avoid contaminations all solvents used should be of HPLC grade or better The plastic materials used for sample purification and matrix preparation should be explicitly recommended for mass spectrometric measurements. Many common plastics may cause polymer contamination of the sample. This will interfere with the subsequent matrix preparation/crystallization and produce polymer signals during spectra acquisition. Sample purification and MALDI target preparation should be performed under clean conditions. Dust and air pollution will decrease the spectra quality. For optimal sample/matrix crystallization on the MALDI target the following environmental conditions are recommended: Operating temperature: 5 - 40°C (41 - 104°F) Operating humidity: 25 – 65 % at 22°C Atmospheric pressure: 75 - 105 kPa For high resolution measurements and low signal to noise values a cleaning of the ion source at regular intervals is absolutely mandatory. This procedure can be performed with the Bruker Daltonics’ source shower target (see ordering information). For a complete cleaning of the ion source please ask Bruker Daltonics’ service team (maldi.support@bdal.de). Page 6 of 19
Recommended sample preparation procedure Binding of samples 1. Re-suspend the Magnetic Beads by shaking for at least 20 times from top to the bottom and reversed or careful vortexing (do not sonificate!) to get a homogenous suspension. Repeat shaking between pipetting steps if necessary. Re-suspend the beads 2. Transfer 20 µl of resuspended Magnetic Beads to a standard thin wall PCR-tube and add 100 µl WB 1. 20 µl beads 100 µl WB1 3. Place the tube in a Magnetic Separator and move the tubes back and forth between adjacent wells 20 times. Wait for 20 seconds to separate the beads from the supernatant. Move back and forth Collect beads 4. Remove the supernatant carefully using a pipette. Avoid contact of pipette tips with the magnetic beads and take care not to remove the beads. Remove supernatant 5. Resuspend the beads in 100 µl WB 1 100 µl WB1 6. Repeat steps 3 and 4 to wash the beads and remove the supernatant Repeat steps 3 and 4 7. Re-suspend the beads in 10 µl BB and add up to 10 µl of the sample. If you have less sample volume adjust to 10 µl by adding deionized water. For the analysis of serum, use 10 - 20 µl serum. Firstly, pipette an equal amount of BB (10 - 20 µl) to 20 µl magnetic beads and subsequently add the serum. 10 µl BB 10 µl sample 8. Incubate the magnetic beads for 1 h at room temperature while swirling the tube from time to time to mix the beads. 1 h incubation Page 7 of 19
Page 1 and 2: CARE products are designed for supp
Page 3 and 4: Table of Contents General Informati
Page 5: Kit Components Materials provided i
Page 9 and 10: 19. Place the tube on the Magnetic
Page 11 and 12: Cleaning of polished steel targets
Page 13 and 14: 8. 1 µl of the turbid suspension h
Page 15 and 16: Analysis of deglycosylated peptides
Page 17 and 18: References 1 Sparbier K, Koch S, Ke
Page 19: Ordering Information Product Size (

Instructions for Use - Glyco Kit MB-LAC AIA - Bruker

Create successful ePaper yourself

Delete template?

Save as template?