Waters MassPREP Hemoglobin Digestion Standard User Manual

[ Care and Use ManUal ]

MassPREP HEMoglobin DigEstion stanDaRD

i. intRoDuction

The MassPREP™ protein digestion standard was prepared by digesting bovine
hemoglobin (SwissProt HBA P01966, HBB P02070) 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
Bovine Hemoglobin 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

Reconstitution

volume (µl)

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

Column i.d.

(mm)

Injection

volume

(µl)

Aver. injected

mass per peptide

(pmole)

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

Part Number

186002325 MassP REP Digestion Standard, Enolase 1

186002326

186002327

186002328

186002329

186002330

186002331

186002333

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.

Product Description Qty per pack

MassPREP Digestion Standard, Phosphorylase b

MassPREP Digestion Standard, Bovine Hemoglobin

MassPREP Digestion Standard, Alcohol Dehydrogenase

MassPREP Digestion Standard, Bovine Serum Albumin

MassPREP Digestion Standard Kit (one of each digestion std.)

MassPREP MALDI Matrix CHCA

MassPREP MALDI Matrix DHB

1

1

1

1

5

5

5

* Calculated for peptide with molecular weight of 1500 g/mol

MassPREP Hemoglobin Digestion Standard 1

[ Care and Use ManUal ]

Sales Offices:

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(Central South Eastern Europe,

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CIS/Russia [phone]

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United Kingdom 44 208 238 6100

©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 715001600 Rev B EY-PDF

MassPREP Hemoglobin Digestion Standard 2

Waters Corporation

34 Maple Street
Milford, MA 01757 U.S.A.
T: 1 508 478 2000
F: 1 508 872 1990
www.waters.com