ETD Verification Kit II

Transcription

1 Instructions for Use ETD Verification Kit II Kit for verifying and tuning electron transfer dissociation (ETD) and proton transfer reaction (PTR) CARE products are designed to support our worldwide customers with highquality consumables, accessories and dedicated kits. The CARE product range is specifically optimized and certified for use with all Bruker Daltonics systems. Language: en Revision B (December 2015)

2 1 Product description 2 2 Storage and stability 2 3 Risk and safety information 3 4 Sample Preparation Procedure 3 5 Manufacturer 5 1 Product description The ETD verification kit II allows the tuning of the electron transfer conditions with a synthetic ETD peptide. This peptide corresponds to a peptide from a tryptic digest of Peptidoglycan recognition protein 2 (from amino acid position ) with the following amino acid sequence: GWHWVGAHTLGHNSR Mono isotopic mass: [M+2H] 2+ : [M+3H] 3+ : [M+4H] 4+ : [M+5H] 5+ : With the (isolated) higher charge states of the ETD peptide with z > 3, the tuning of the proton transfer reaction (PTR) can be done as well. Ordering Information Product ETD verification kit II ETD consumable kit ETD II consumable kit ETD III consumable kit Part No. 2 Storage and stability The tube box is shipped at ambient temperatures. We recommend storing the proteins on arrival at less than 0 C. Peptide solutions should be aliquoted and frozen. We do not recommend refreezing samples after thawing. Page 2 of 5 ETD Verification Kit II IFU Revision B

3 3 Risk and safety information The ETD Verification Kit II is not classified according to Regulation (EC) No. 1272/2008 and is therefore not classified according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS). In the manufacturer's experience, the product has no harmful effect when used and handled according to specifications. However, all materials may present unknown hazards and should be handled with caution. For more information, read the Material Safety Data Sheet available for download at Additional chemicals may be required for procedures described in these Instructions for Use. Carefully read the Material Safety Data Sheet provided by the supplier and follow general safety regulations when handling chemicals or biohazardous material. 4 Sample Preparation Procedure Preliminary remarks Poor sample preparation will degrade sensitivity and result in poor reproducibility. For best results use only chemicals of highest available purity. Chemicals and materials required HPLC grade acetonitrile Syringe, 250 µl HPLC grade water Syringe pump Formic acid, e.g. Merck # Connector from syringe to MS instruments Acetonitrile is classified as hazardous chemical. DANGER: H: 225, 319, 302, 312, 332; P: 210, 280, Formic acid is classified as hazardous chemical. DANGER: H: 314; P: 280, Preparation of peptide working solution Preparation of solvent: 0.1% formic acid in water/acetonitrile, 1:1 (v/v) Preparation of peptide stock solution: thoroughly dissolve content of one vial in 100 μl solvent (vortexing is recommended) Preparation of peptide working solution: mix 900 μl solvent μl peptide stock solution. ETD and PTR verification measurements For these measurements, the direct infusion of the peptide working solution is recommended. For further details of tuning ETD and PTR conditions, please refer to the tutorial 'ETD and PTR with the HCTultra'. Revision B ETD Verification Kit II IFU Page 3 of 5

4 Results of ETD MS/MS measurements using the ETD verification kit Figure 1 shows a typical ETD MS/MS spectrum (precursor ion Th, [M+3H]3+) obtained from the ETD verification kit. Measurement was performed on a Bruker Daltonics HCTultra PTM discovery system. Table 1 lists the theoretical m/z values for the observed fragment ions. Figure 1 ETD MS/MS spectrum of ETD verification kit Fragment ion Monoisotopic mass (singly charged) Fragment ion Monoisotopic mass (singly charged) (z+1) (z+1) (z+1) (z+1) c c (z+1) (z+1) (z+1) c c c (z+1) (z+1) c c c (z+1) (z+1) c c c (z+1) (z+1) c Table 1 Theoretical m/z values of typical fragment ions Page 4 of 5 ETD Verification Kit II IFU Revision B

5 5 Manufacturer Bruker Daltonik GmbH Fahrenheitstraße Bremen Germany Support esi.support@bruker.com Phone: +49 (421) Fax: +49 (421) Sales Information care@bruker.com Phone: +49 (421) Web: For research use only. Not for use in diagnostic procedures. # Descriptions and specifications supersede all previous information and are subject to change without notice. Copyright 2015 Bruker Daltonik GmbH Revision B ETD Verification Kit II IFU Page 5 of 5