Sartorius stedim Sartobind S User manual

Sartobind®S Demo Kit
Purification of a protein with Sartobind S Membrane Adsorbers
85034-535-14
1. Storage before use
The kit solutions have been filtered
sterile (0.2 μm) but no preservatives
have been added to the buffers. Thus
the kit should be kept refrigerated at
+4 to +8°C after opening bottles.
2. Introduction
The kit has been developed to
demonstrate the capability of
Sartobind Membrane Adsorbers to
purify quickly and easily a protein
(cytochrome c) from a contaminant
(Amido Black).
A trained person can demonstrate
the purification in one minute.
A beginner can perform the purifica-
tion within five minutes.
3. Kit contents
Sartobind S 15 1
Equilibration
buffer 2 +50 ml
Loading solution 50 ml
Elution buffer 50 ml
10 ml syringe 1
Order code 90-KIT-01
4. Technical data Sartobind S 15
Membrane volume 0.41 ml
Bed height 0.8 mm
Number of layers 3
Membrane
diameter 25 mm
Dimensions 36 +28 mm
Ligand Sulfonic acid
Degree of
substitution 2–5 μeq/cm2
Binding capacity 0.8 mg/cm2
29 mg/ml
lysozyme
Membrane matrix Cross-linked rein-
forced cellulose
Nom. Pore size >3 μm
Membrane
thickness 275 μm
Spacer No spacer
Membrane area 15 cm2
Housing Polysulfone
Connector inlet Female Luer
outlet Male Luer
Flow at 0.1 MPa
(1 bar, 14.5 psi) 130 ml/min
Recommended
flow rate 10–150 ml/min
Maximum 0.6 MPa
pressure (6 bar, 87 psi)
Chemical All commonly used
stability chromatography
buffers
pH stability
Long term 2–12
Short term 1–14
Storage in equilibration
buffer containing
20% ethanol
4. Operation of the units
We recommend to use the enclosed
syringe for the demonstration:
Use recommended volumes as listed
in table.
Buffers, protein solution and volumes
Equilibration 10 mM potassium
buffer: 10 ml phosphate buffer
pH 7.0
Loading solution: Cytochrome c
5 ml (2 mg/ml) in
equilibration
buffer + Amido
Black (5 μg/ml)
Washing buffer: Equal to
5–10 ml equilibration buffer
Elution buffer: Equilibration
3 ml buffer + 1 M NaCl
The buffers and protein solution can
be prepared by yourself to repeat the
demonstration. S 15 unit is reusable.
4.1 Equilibration
Hold the unit in a vertical position.
Remove the inlet cap. Fill 10 ml
syringe with 10 ml equilibration
buffer and connect to the inlet of the
unit. Remove the outlet cap. Fill the
unit and remove any remaining air by
moving the plunger up and down
with short strokes. Make sure that
the unit is completely filled with the
fluid.
Important note:
The unit has to be filled completely
with fluid throughout the operation
to ensure even flow and distribution
of the feed stream.
Outlet
Inlet
Flow
direction

4.2 Loading start
Push approximately 5 ml of protein
solvent through the unit at any
speed. If you detect air bubbles just
lift plunger a few mm to let it ascend
into the syringe.
4.2 Loading end
The red protein is bound on the
membrane and the contaminant
Amido Black flows through.
4.3 Washing
Wash with 5–10 ml of equilibration
buffer.
4.4 Elution
Elute with 3 ml elution buffer.
The unit may not run dry during the
operation.
The red protein is eluted and concen-
trated at the same time.
4.5 Regeneration
Regenerate the unit by application of
10 ml of equilibration buffer.
4.6 Storage after use
Keep the used unit filled with equili-
bration buffer in presence of an
antimicrobial preservative such as
20% ethanol.
4.7 Stability
The membranes are stable in a pH
range of 2–12 (long term) and 1–14
(short term). Do not use oxidative
agents, such as hypochlorite or H2O2.
4.8 Use of peristaltic pumps or
liquid chromatography systems
For the operation of Membrane
Adsorber units with a peristaltic
pump or a LC system, specific Luer
Lock adapters are needed (see Order-
ing Information). Proceed as
described under 4.1 Equilibration.
When the unit is filled completely
with equilibration buffer, close the
outlet and remove the syringe. Start
your LC system or peristaltic pump
at a very low flow rate. When fluid
emerges, stop the pump, connect the
tubing via a Luer Lock adapter to the
inlet of the unit. Make sure that no
air is introduced. Remove the lower
cap. Run the pump until fluid
emerges from the outlet of the unit
and stop it. Then connect outlet of
the unit via Luer Lock adapter to the
LC detector and proceed with load-
ing.
Important note
For in vitro use only.
Use the units only as described in this
operating instructions.
Follow safety regulations: Wear
protective gloves, safety glasses and
a lab coat during work.
Related products for
laboratory scale
Order number Description
S5f Sartobind S 5,
5 cm2membrane
area, single layer,
15 pcs. Single use
S15X Sartobind S 15,
15 cm2, 3 layers,
2 pcs., reusable
S75X Sartobind S 75,
75 cm2, 15 layers,
2 pcs., reusable
S100X Sartobind S 75,
100 cm2, 5 layers,
2 pcs., reusable
Sartorius Stedim Biotech GmbH
August-Spindler-Strasse 11
37079 Goettingen, Germany
Phone [phone]
Fax [phone]
www.sartorius-stedim.com
USA Toll-Free +1.800.368.7178
UK +44.1372.737159
France +33.442.845600
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Spain +34.91.3586102
Japan +81.3.3740.5407
Sartobind®is a trademark of
Sartorius Stedim Biotech GmbH,
Goettingen, Germany.
Technical data subject to change
without notice.
Printed and copyrighted
by Sartorius · W404.18
Publication No.: SL-6139-e08041