Millipore Centriprep 50k, User Manual

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Chemical Compatibility
Centriprep® devices are intended for use with biological fluids and aqueous solutions. Before use, check
the sample for chemical compatibility with the device. The solutions listed in the following table have been
evaluated for chemical compatibility with Centriprep® devices.
Acids Concentration Concentration
Acetic acid ≤ 80% Phosphoric acid ≤ 30%
Formic acid ≤ 50% Sulfuric acid ≤ 3%
Hydrochloric acid ≤ 1.0 N Trichloroacetic acid (TCA) ≤ 10%
Lactic acid ≤ 50% Trifluoroacetic acid (TFA) ≤ 50%
Nitric acid ≤ 10%
Alkalis
Ammonium hydroxide ≤ 5% Sodium hydroxide ≤ 0.1 N
Alcohols
n-Butanol ≤ 90% Isopropanol ≤ 70%
Ethanol ≤ 70% Methanol ≤ 60%
Detergents
Alconox® detergent ≤ 1% Sodium deoxycholate ≤ 5%
Lubrol® PX detergent ≤ 0.1% Tergazyme® detergent ≤ 1%
Nonidet™ P-40 surfactant ≤ 2% Triton® X-100 surfactant ≤ 0.002%
PyroCLEAN ™ detergent 1:10 Tween® 20 surfactant ≤ 0.1%
Sodium dodecyl sulfate (SDS) ≤ 2%
Organic solvents
Acetone not recommended Dioxane not recommended
Acetonitrile ≤ 30% Ethyl acetate not recommended
Benzene not recommended Formaldehyde ≤ 5%
Carbon tetrachloride not recommended Pyridine not recommended
Chloroform not recommended Tetrahydrofuran not recommended
Dimethyl formamide ≤ 50% Toluene not recommended
Dimethyl sulfoxide (DMSO) ≤ 5%
Miscellaneous
Ammonium sulfate Saturated Phenol ≤ 1%
Diethyl pyrocarbonate ≤ 0.2% Phosphate buffer (pH 8.2) ≤ 1 M
Dithiothreitol (DTT) ≤ 0.1 M Polyethylene glycol ≤ 10%
Glycerine ≤ 70% Sodium carbonate ≤ 20%
Guanidine HCl ≤ 6 M Sodium thiocyanate ≤ 3 M
Guanidine thiocyanate ≤ 0.5 M Tris buffer (pH 8.2) ≤ 1 M
Imidazole ≤ 100 mM Urea ≤ 8 M
Mercaptoethanol ≤ 0.1 M
Product Ordering Information
This section lists the catalogue numbers for Centriprep® Ultrafiltration Devices. See the Technical
Assistance section for contact information. You can purchase these products on-line at
www.millipore.com/products.
Description 24/pk 96/pk
For in vitro diagnostic use
Centriprep® 10K device (10,000 MWCO) 4304 4305
Centriprep® 30K device (30,000 MWCO) 4306 4307
For research use only
Centriprep® 3K device (3,000 MWCO) 4302 4303
Centriprep® 50K device (50,000 MWCO) 4310 4311
Notice
The information in this document is subject to change without notice and should not be construed as a
commitment by Merck Millipore Ltd. (“Millipore”) or an affiliate. Neither Merck Millipore Ltd. nor any of its
affiliates assumes responsibility for any errors that may appear in this document.
Technical Assistance
For more information, contact the office nearest you. Up-to-date world-wide contact information is
available on our web site at www.millipore.com/offices. You can also visit the tech service page on our web
site at www.millipore.com/techservice.
Standard Warranty
The applicable warranty for the products listed in this publication may be found at
www.millipore.com/terms (within the “Terms and Conditions of Sale” applicable to your purchase
transaction).
The M logo is a trademark of Merck KGaA, Darmstadt, Germany.
Millipore, Centriprep, and Ultracel are registered trademarks of Merck KGaA.
All trademarks of third parties are the property of their respective owners.
© 2013 EMD Millipore Corporation. Billerica, MA, U.S.A. All rights reserved.
PR04321, Rev. A, 09/13
Made in Ireland
Merck Millipore Ltd.
Tullagreen,
Carrigtwohill,
Co. Cork, IRL
Performance Figures
Typical Filtration Profiles for Centriprep® Devices Using Common Dilute Protein Solutions
Centriprep® 3K
Device
Centrifugation Time (min)
Concentr
at
e V
olume (mL)
Operating
Parameters:
● g-force: 3,000 × g
● temperature: 25 °C
● swinging-bucket rotor
● solute: 0.25 mg/mL
Cytochrome c
5, 10, and 15 mL
starting volumes
Filtrate decanted
at times indicated
Centriprep® 10K
Device
Centrifugation Time (min)
Concentr
at
e V
olume (mL)
Operating
Parameters:
● g-force: 3,000 × g
● temperature: 25 °C
● swinging-bucket rotor
● solute: 1 mg/mL
Bovine serum albumin
5, 10, and 15 mL
starting volumes
Filtrate decanted
at times indicated
Centriprep® 30K
Device
Centrifugation Time (min)
Concentr
at
e V
olume (mL)
Operating
Parameters:
● g-force: 1,500 × g
● temperature: 25 °C
● swinging-bucket rotor
● solute: 1 mg/mL
Bovine serum albumin
5, 10, and 15 mL
starting volumes
Filtrate decanted
at times indicated
Centriprep® 50K
Device
Centrifugation Time (min)
Concentr
at
e V
olume (mL)
Operating
Parameters:
● g-force: 1,500 × g
● temperature: 25 °C
● swinging-bucket rotor
● solute: 1 mg/mL
Bovine serum albumin
5, 10, and 15 mL
starting volumes
Filtrate decanted
at times indicated
How to Quantify Recoveries
Calculate total recovery, percent concentrate recovery, and percent filtrate recovery using the method
below. This procedure provides a close approximation of recoveries for solutions having concentrations up
to roughly 20 mg/mL.
NOTE: Appropriate assay techniques include absorption spectrophotometry, radioimmunoassay, refractive
index, and conductivity.
Weighing Procedure
The density of most dilute proteins is nearly equal to the density of water (i.e., 1 g/mL). Using this property,
the concentrate and filtrate volumes can be quantified by weighing them and converting the units from
grams to milliliters. This technique is valid only for solutions with concentrations of approximately
20 mg/mL or less.
1. Before use, weigh the sample container and an empty test tube (15 mL capacity).
2. Fill the sample container with solution and reweigh.
3. Assemble the centrifugal filter device and spin per instructions. Decant filtrate into preweighed
test tube. If further concentration is desired, spin a second time, decanting filtrate into test tube
containing first filtrate. Reweigh the test tube plus filtrate(s).
4. Disassemble the device and reweigh the sample container plus concentrate.
5. Subtract weight of empty test tube or sample container to calculate the weights of the starting
material, filtrate, and concentrate.
6. Assay the starting material, filtrate, and concentrate to determine solute concentration.
7. Calculate recoveries using the weight/volume data and the measured concentrations as follows:
% concentrate recovery = 100 ×
W
c
× C
c
W
o
× C
o
% filtrate recovery = 100 ×
W
f
× C
f
W
o
× C
o
%
total recovery = % concentrate re % filtrate recovery
W
c
= total weight of concentrate before assay
W
o
= weight of original starting material
W
f
= weight of filtrate
C
c
= concentrate concentration
C
o
= original starting material concentration
C
f
= filtrate concentration
Specifications
Sample container capacity
15 mL
Final concentrate volume at 25 °C 0.5–0.6 mL
Filtrate collector capacity 12 mL
Maximum relative centrifugal force
Centriprep® 3K device 3,000 × g
Centriprep® 50K device 1,500 × g
Active membrane area
2.84 cm 2
Hold-up volume (membrane and support) 0.5 mL
Dimensions
Diameter of twist-lock cap 36.5 mm (cap resides outside centrifuge carrier)
Diameter of sample container 28.1 mm
Length 135 mm
Materials of Construction
Membrane Ultracel ® regenerated cellulose
Sample container Polycarbonate
Membrane support Polycarbonate
Filtrate collector Polycarbonate
Twist-lock cap Nylon
Air seal cap Polypropylene
PR04321_RevA.indd 2 8/29/2013 1:35:01 PM