Instructions for Use ETD Verification Kit Kit for verifying and tuning electron transfer dissociation (ETD) 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. www.bruker.com/care Language: en Revision B (December 2015)
1 Product description 2 2 Storage and stability 2 3 Risk and safety information 3 4 Sample Preparation Procedure (Recommendation) 3 5 Manufacturer 5 1 Product description The ETD verification kit allows the tuning of the electron transfer conditions using Substance P peptide. Amino acid sequence of Substance P: RPKPQQFFGLM, C-terminal amidated. Mono isotopic mass: [M+2H] 2 +: 674.82 [M+3H] 3 +: 450.21 Ordering Information Product Part No. ETD verification kit 8243460 ETD verification kit II 8255659 ETD consumable kit 8243459 ETD II consumable kit 8255915 ETD III consumable kit 8250942 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 IFU Revision B
3 Risk and safety information Caution substances are not yet fully tested (EU). The kit does not have to be labeled according to Regulation (EC) No 1272/2008. When used and handled according to specifications, the product does not have any harmful effects according to our experience. All materials may present unknown hazards and should be used with caution. Observe the general safety regulations when handling chemicals. 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 (Recommendation) 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 # 100264 Connector from syringe to MS instruments Acetonitrile is classified as hazardous chemical. DANGER: H: 225, 319, 302, 312, 332; P: 210, 280, 305+351+338 Formic acid is classified as hazardous chemical. DANGER: H: 314; P: 280, 305+351+338 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 + 100 μl peptide stock solution. ETD verification measurements Use the peptide working solution for ETD verification as described in the instrument manual. Revision B ETD Verification Kit IFU Page 3 of 5
Results of ETD MS/MS measurements using the ETD verification kit Figure 1 shows a typical ETD MS/MS spectrum (precursor ion 450.21 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 Table 1 ETD MS/MS spectrum of ETD verification kit Theoretical m/z values of typical fragment ions Fragment ion Monoisotopic mass (singly charged) c2 271.19 c4 496.34 c5 624.39 c6 752.45 c7 899.52 Monoisotopic mass (singly charged) c8 1046.59 523.80 c9 1103.61 552.32 c10 1216.69 608.85 (z+1) 2 246.12 (z+1) 5 597.27 (z+1) 6 725.33 (z+1) 7 853.39 (z+1) 9 1078.56 Page 4 of 5 ETD Verification Kit IFU Revision B
5 Manufacturer Bruker Daltonik GmbH Fahrenheitstraße 4 28359 Bremen Germany Support E-mail: esi.support@bruker.com Phone: +49 (421) 2205-420 Fax: +49 (421) 2205-106 Sales Information E-mail: care@bruker.com Phone: +49 (421) 2205-0 Web: www.bruker.com/care For research use only. Not for use in diagnostic procedures. # 8243460 Descriptions and specifications supersede all previous information and are subject to change without notice. Copyright 2015 Bruker Daltonik GmbH Revision B ETD Verification Kit IFU Page 5 of 5