Waters MassPREP Phosphorylase B Digestion Standard - Manual

[ Care and Use ManUal ]
MassPREP Hemoglobin Digestion Standard
MassPREP PhosPhoRylasE b DigEstion stanDaRD
i. intRoDuction
The MassPREP protein digestion standard was prepared by digesting Rabbit
Phosphorylase b (SwissProt P00489) with sequencing grade Trypsin. The sample
was reduced and alkylated (iodoacetamide) prior to digestion. The resulting
digest does not contain any undigested protein, Trypsin, alkylation/reduction
agents, or very hydrophilic components. Each batch of Phosphorylase b diges-
tion 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. % trifluo-
roacetic 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 nmole of pro-
tein. For HPLC column evaluation, the recommended sample dilution is 00
μl of 0. % trifluoroacetic acid in water, or 0.- % formic acid in water.
Injection volumes and mass loads are shown in Table .
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
. Dissolve protein digestion standard in .0 mL of 0.% trifluoroacetic
acid/0 % acetonitrile (v/v). The concentration of peptides in this stock
solution is ~ 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 80 μl of deionized water. The
concentration of this working solution is 00 fmol/μl.
4. Mix μl of standard with μl of MALDI matrix (high quality MassPREP
MALDI Matrix is recommended, see table below). Deposit 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 90.200(c) and
Appendix A and B, and are thus exempt from the requirements of the Hazard
Communication Standard, including the requirement to supply MSDSs.
Reconstitution
volume (µl)
Column i.d.
(mm)
Injection
volume
(µl)
Aver. injected
mass per peptide
(pmole)
00 4.6 75.0 750.0
00 2. 5.6 56.3
00 .0 3.5 35.4
00 0.32 0.4 3.6
00 0.8 0. .
00 0.075 0.02 0.2
Part Number Product Description Qty per pack
86002325 MassPREP Digestion Standard, Enolase
86002326 MassPREP Digestion Standard, Phosphorylase b
86002327 MassPREP Digestion Standard, Bovine Hemoglobin
86002328 MassPREP Digestion Standard, Alcohol Dehydrogenase
86002329 MassPREP Digestion Standard, Bovine Serum Albumin
86002330 MassPREP Digestion Standard Kit (one of each digestion std.) 5
8600233 MassPREP MALDI Matrix CHCA 5
86002333 MassPREP MALDI Matrix DHB 5