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/cm
2
Binding capacity 0.8 mg/cm
2
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 cm
2
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
Amido
Black (5 μg/ml)
Washing buffer: Equal to
5 –10 ml
equilibration buffer
Elution buffer:
Equilibration
3 ml
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
