[ Care and Use Manual ]
MassPREP ADH Digestion Standard
IV. Reconstitution of MassPREP protein digestion
standard for MALDI MS
I.Introduction
The MassPREP™ protein digestion standard was prepared by digesting yeast
1.
alcohol dehydrogenase (ADH, SwissProt P00330) with sequencing grade
Dissolve protein digestion standard in 1.0 mL of 0.1% trifluoroacetic
Trypsin. The sample was reduced and alkylated (iodoacetamide) prior to diges-
acid/10 % acetonitrile (v/v). The concentration of peptides in this stock
tion. The resulting digest does not contain any undigested protein, Trypsin,
solution is ~1 pmol/μl.
alkylation/reduction agents, or very hydrophilic components. Each batch of
2. Dispense 20 μl aliquots in 0.5 ml eppendorf tube and store at -20 °C
ADH digestion standard is prepared under strict quality control criteria con-
prior to use.
firmed on the included certificate of analysis. Protein digestion standards may
3. Thaw the content of a tube and add 180 μl of deionized water. The
be used for a variety of applications, including evaluation of HPLC columns,
concentration of this working solution is 100 fmol/μl.
evaluation of mass spectrometry and chromatography instrumentation performance, mass spectrometer calibration, or as an HPLC standard.
4. Mix 1 μl of standard with 1 μl of MALDI matrix (high quality MassPREP
MALDI Matrix is recommended, see table below). Deposit 1 μl on the
II. storage and stability
MALDI target for analysis.
The lyophilized powder is stable at room temperature until the expiration
V. Ordering information
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
Part Number
Product Description
8 °C. Stability can be extended by freezing samples at -20 °C.
186002325
MassPREP Digestion Standard, Enolase
1
186002326
MassPREP Digestion Standard, Phosphorylase b
1
Note: The contamination of sample with endo- and exopeptidase enzymes may
186002327
MassPREP Digestion Standard, Bovine Hemoglobin
1
cause rapid sample degradation.
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
III. Reconstitution of MassPREP protein digestion
standard for HPLC
Qty per pack
The protein digestion standards were prepared by digesting 1 nmole of
Note: Each MassPREP Digestion standard is sufficient for a thousand or more
protein. For HPLC column evaluation, the recommended sample dilution is
MALDI MS analyses.
100 μl of 0.1% trifluoroacetic acid in water, or 0.1-1% formic acid in water.
MSDS information: Waters Digestion Standards contain various non hazard-
Injection volumes and mass loads are shown in Table 1.
ous peptides in powder form. They are for laboratory use only; solely for
Table 1: Recommended Mass Load for HPLC Columns Evaluation
Reconstitution
volume (µl)
Column i.d.
(mm)
100
100
100
100
100
100
4.6
2.1
1.0
0.32
0.18
0.075
Injection
volume
(µl)
75.0
15.6
3.5
0.4
0.1
0.02
purposes of scientific experimentation, analysis, research or development.
Waters Digestion Standards should be used only as directed and in accor-
Aver. injected
mass per peptide
(pmole)
750.0
156.3
35.4
3.6
1.1
0.2
dance with good laboratory 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.
* Calculated for peptide with molecular weight of 1500 g/mol
MassPREP ADH Digestion Standard
[ Care and Use Manual ]
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©2007 Waters Corporation. Waters, The Science of
What’s Possible, and MassPREP are trademarks of
Waters Corporation.
Printed in the U.S.A.
July 2007 715001593 Rev B EY-PDF
MassPREP ADH Digestion Standard
Waters Corporation
34 Maple Street
Milford, MA 01757 U.S.A.
T: 1 508 478 2000
F: 1 508 872 1990
www.waters.com