Waters MassPREP Enolase Digestion Standard - Manual

[ Care and Use ManUal ]
MassPREP Enolase Digestion Standard 1
I. IntroductIon
The MassPREP ™ protein digestion standard was prepared by digesting Yeast
Enolase (SwissProt P00924) with sequencing grade Trypsin. The resulting
digest does not contain any undigested protein, Trypsin, and very hydrophilic
components. Each batch of Enolase digestion standard is prepared under strict
quality control criteria confirmed on the included certificate of analysis.
Protein digestion standards may be used for a variety of applications,
including evaluation of HPLC columns, evaluation of mass spectrometry and
chromatography instrumentation performance, mass spectrometer calibration,
or as an HPLC standard.
II. storage and stabIlIty
The lyophilized powder is stable at room temperature until the expiration
date printed on the label. Once reconstituted in high purity water or in 0.1 %
trifluoroacetic acid in water, the solution is stable for one week when stored at
8 °C. Stability can be extended by freezing samples at -20 °C.
Note: The contamination of sample with endo- and exopeptidase enzymes may
cause rapid sample degradation.
III. reconstItutIon of MassPreP ProteIn dIgestIon
standard for HPlc
The protein digestion standards were prepared by digesting 1 nmole of pro-
tein. For HPLC column evaluation, the recommended sample dilution is 100
μl of 0.1 % trifluoroacetic acid in water, or 0.1-1 % formic acid in water.
Injection volumes and mass loads are shown in Table 1.
Table 1: Recommended mass load for HPLC columns evaluation
* Calculated for peptide with molecular weight of 1500 g/mol
Iv. reconstItutIon of MassPreP ProteIn dIgestIon
standard for MaldI Ms
1. Dissolve protein digestion standard in 1.0 mL of 0.1% trifluoroacetic
acid/10 % acetonitrile (v/v). The concentration of peptides in this stock
solution is ~1 pmol/μl.
2. Dispense 20 μl aliquots in 0.5 ml eppendorf tube and store at -20 °C
prior to use.
3. Thaw the content of a tube and add 180 μl of deionized water. The
concentration of this working solution is 100 fmol/μl.
4. Mix 1μl of standard with 1μl of MALDI matrix (high quality MassPREP
MALDI Matrix is recommended, see table below). Deposit 1 μl on the
MALDI target for analysis.
v. orderIng InforMatIon
Note: Each MassPREP Digestion standard is sufficient for a thousand or more
MALDI MS analyses.
MSDS information: Waters Digestion Standards contain various non hazardous
peptides in powder form. They are for laboratory use only; solely for purposes of
scientific experimentation, analysis, research or development. Waters Digestion
Standards should be used only as directed and in accordance with good labora-
tory practice. All Waters Digestion Standards are non hazardous as defined
by OSHA’s Hazard Communication Standard, 29 CFR 1910.1200(c) and
Appendix A and B, and are thus exempt from the requirements of the Hazard
Communication Standard, including the requirement to supply MSDSs.
MassPreP enolase dIgestIon standard
Reconstitution
volume
(µl)
Column i.d.
(mm)
Injection volume
(µl)
Aver. injected mass
per peptide
(pmole)
100 4.6 75.0 750.0
100 2.1 15.6 156.3
100 1.0 3.5 35.4
100 0.32 0.4 3.6
100 0.18 0.1 1.1
100 0.075 0.02 0.2
Part Number Product Description Qty pe
pack
186002325 MassPREP Digestion Standard, Enolase 1
186002326 MassPREP Digestion Standard, Phosphorylase b 1
186002327 MassPREP Digestion Standard, Bovine Hemoglobin 1
186002328 MassPREP Digestion Standard, Alcohol Dehydrogenase 1
186002329 MassPREP Digestion Standard, Bovine Serum Albumin 1
186002330 MassPREP Digestion Standard Kit (one of each digestion std.) 5
186002331 MassPREP MALDI Matrix CHCA 5
186002333 MassPREP MALDI Matrix DHB 5