Instructions for Use - Glyco Kit MB-LAC Con A - Bruker

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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 1 (for glyco-proteins) or BB 2 (for glyco-peptides) 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 1 or BB 2 (10 - 20 µl) to 20 µl magnetic beads and subsequently add the serum. 10 µl BB1 or BB2 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 Washing of beads 9. Place the tube on the Magnetic Separator and wait for 20 seconds to separate the beads from the supernatant. (Note the movement of the magnetic beads in the tube.) Collect beads 10. Remove the supernatant carefully. Avoid contact of pipette tips with the magnetic beads and take care not to remove the beads. Remove supernatant 11. Add 100 µl WB 1.1 (for glyco-proteins) or WB 2.1 (for glyco-peptides). 100 µl WB1.1 or WB2.1 12. Move the tubes back and forth between adjacent wells at least 20 times to wash the beads. Move back and forth 13. Wait 20 seconds for collecting magnetic beads at the wall of the tubes and remove the supernatant carefully. Collect beads Page 8 of 21
14. Transfer the tube from the Magnetic Separator to an alternative device. Add 100 µl WB2 and place the tube back on the Magnetic Separator. Move the tubes back and forth between adjacent wells 20 times. 100 µl WB2 Move back and forth 15. Wait 20 seconds for collecting magnetic beads at the wall of the tubes and remove the supernatant carefully. Collect beads Remove supernatant 16. Repeat steps 14 and 15 once. Repeat steps 14 and 15 Elution of isolated peptides/proteins 17. For the elution of captured peptides and proteins add 10 µl EB and mix thoroughly. 10 µl EB 18. Incubate for 25 minutes while mixing from time to time. 25 min incubation 19. Place the tube on the Magnetic Separator and wait for 30 seconds for collecting the beads at the wall of the tube. Collect beads 20. Finally, transfer the elution solution containing purified glycopeptides/-proteins into a fresh tube. Transfer eluate Page 9 of 21
Page 1 and 2: CARE products are designed for supp
Page 3 and 4: Table of Contents General Informati
Page 5 and 6: Kit Components Materials provided i
Page 7: Recommended sample preparation proc
Page 11 and 12: MALDI-TOF MS Target Preparation MAL
Page 13 and 14: Matrix preparation on the MALDI-MS
Page 15 and 16: 7. After drying of the spots the ta
Page 17 and 18: Examples Isolation of a small glyco
Page 19 and 20: References 1 Sparbier K, Koch S, Ke
Page 21: Ordering Information Product Size (

Instructions for Use - Glyco Kit MB-LAC Con A - Bruker

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