Waters MassPREP Hemoglobin Digestion Standard User Manual

Care and use manual, Massprep hemoglobin digestion standard, I. introduction

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[ Care and Use ManUal ]

MassPREP Hemoglobin Digestion Standard



MassPREP HEMoglobin DigEstion stanDaRD

i. intRoDuction

The MassPREP

protein digestion standard was prepared by digesting bovine

hemoglobin (SwissProt HBA P0966, HBB P02070) with sequencing grade
Trypsin. The sample was reduced and alkylated (iodoacetamide) prior to diges-
tion. 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 stan-
dards 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. %
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  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

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