MRMplus Retention Time Marker

MRMplus Retention Time Marker
Cat. No: FNK-FMR-002


Description
MRMplus Retention Time Marker is the mixture of marker peptides (12 synthetic peptides) for calculating the retention times of LC in LC-MS/MS system for scheduled-MRM. The segment value of each MRMplus Standard Peptide is easily converted by using this marker and the conversion software.
 

Component
  * Dilute 1 µl of Stock Solution into 199 µl of sample diluent for your LC system as Working Solution. When you use TFA as the sample diluent, grass ampule type product (eg. #28902 from Thermo Scientific) is recommended. Working Solution should be stored at 4ºC

NOTE: AVOID FREEZE-THAW CYCLES for the stock solution

NOTE: Please use working solution within ONE WEEK



General Protocol

* This protocol is optimized for QTRAP5500 System (nanoESI, ABSciex). For the other LC-MS/MS systems, optimal condition should be determined by users.

1. Inject 0.2 µl of the Working Solution into LC.
2. Optimize LC conditions and injection volume.
3. Input the values of Q1 and Q3 for 12 marker peptides and conduct MRM analysis without scheduling function.
4. Write down the detected retention time of 12 marker peptides into spread sheet.
5. Convert segment values of internal standard peptides specific to your target proteins into expected retention time using the detected retention time and conversion software (RTimeToSegment.exe).
6. Input the expected retention time into your LC-MS/MS system.
7. Mix 1 µl Retention Time Marker for 1 µg sample, and then add the suitable amount of internal standard peptide. Dilute the mixture to 10 µl with sample diluent, use 2 µl for conduct scheduled MRM analysis.

NOTE: You can find the information of the Q1, Q3 values, and conversion software (RTimeToSegment.exe) on FUNAKOSHI website (http://www.funakoshi.co.jp/contents/7979).

NOTE: You should set a wide range of expected retention time for scheduling, or you would miss the target peptide.

NOTE: Some marker peptides may appear closely each other or sometimes interchanged depending on your LC conditions, such as gradient, colums used, or room temperature.