Waters MassPREP Protein Standard Mix - Manual (Page 2)

[ CARE AND USE MANUAL ]
MassPREP Protein Standard Mix 2
II. STORAGE AND STABILITY
Upon receiving the sample, store immediately at -20 °C. The lyophilized
powder is stable at -20 °C until the expiration date printed on the label.
Once reconstituted, the solution should be used immediately for best
performance. Stability can be extended by freezing samples at -20 °C.
III. RECONSTITUTION OF MASSPREP PROTEIN STANDARD MIX
FOR HPLC/UPLC USE
Each vial of the MassPREP Protein Standard Mix contains approximately
123 µg of total protein. Table 2 outlines the individual concentra-
tions. For ACQUITY UPLC BEH C
4
1.7 μm, 2.1 x 50 mm column evalu-
ation (P/N 186004495) the recommended reconstitution volume is
100 μL of 0.1% trifluoroacetic acid in water with a 5 µL load to make
a sample containing 1228 ng/ µL of total protein.
Table 2: Amount of Each Protein per Vial
Protein Amount (µg)
Ribonuclease A 4.15
Cytochrome c 5.65
Bovine Serum Albumin 20.15
Myoglobin 12.50
Enolase 21.45
Phosphorylase b 58.90
MSDS information: Waters Protein and Peptide 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 Protein and Peptide Standards
should be used only as directed and in accordance 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.
AU
0.00
0.15
0.30
12 24 36 48 min
1
2
3
4
5
6
0
0.00
0.04
0.08
0.12
0.16
AU
0.20
0.24
2 4 6 8 10 12 14 16 18 20 22 24 Min
Column: ACQUITY UPLC ® BEH300 C
4
, 1.7 µm, 2.1 x 150 mm
Mobile Phase A: 0.1% TFA in water
Mobile Phase B: 0.1% TFA in 100% acetonitrile
Flow Rate:
0.2 mL/min
Gradient: 20-55% B in 60 min
Column Temp.: 40 ºC
Injection:
3.3 µL
Detection λ: 220 nm
Column: ACQUITY UPLC BEH C
4
, 1.7 µm, 2.1 x 50 mm
Mobile Phase A: 0.1% TFA in Water
Mobile Phase B: 0.08% TFA in Acetonitrile
Flow Rate: 0.2 mL/min
Gradient: 22-90% B in 16 min
Column Temp: 40 °C
Injection: 5 µL of the MassPREP Protein Standard for a load of
0.615 µg of total protein
Detection λ: 220 nm
Sample: MassPREP Protein Standard mixture prepared in 0.1% TFA in 5% acetonitrile
1. Ribonuclease A (0.04 mg/mL)
2. Cytochrome c (0.06 mg/mL)
3. Bovine Serum Albumin (0.20 mg/mL)
4. Myoglobin (0.13 mg/mL)
5. Enolase (0.22 mg/mL)
6. Phosphorylase b (0.59 mg/mL)
Sample: MassPREP Protein Standard mixture prepared in 0.1% TFA in 5% acetonitrile
1. Ribonuclease A (0.04 mg/mL)
2. Cytochrome c (0.06 mg/mL)
3. Bovine Serum Albumin (0.20 mg/mL)
4. Myoglobin (0.13 mg/mL)
5. Enolase (0.22 mg/mL)
6. Phosphorylase b (0.59 mg/mL)
Figure 1: MassPREP Protein Standard Mix on an
ACQUITY UPLC BEH C4 1.7 μm 2.1 x 150 mm
Figure 2: MassPREP Protein Standard Mix on an
ACQUITY UPLC BEH C4, 1.7 μm, 2.1 x 50 mm