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71-5002-34 AB 2002-08 • p1
inst
ructions
SOURCE 15PHE PE 4.6/100
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SOURCE
Quick Information
SOURCE 15PHE PE 4.6/100 is a pre-packed PEEK column for rapid
preparative hydrophobic interaction chromatography of proteins and
oligonucleotides.
Read the Instruction
The instructions on this page will help you get started quickly with
your new column.
The other side gives more in-depth information on optimisation and
trouble-shooting.
Column data
Matrix:
Polystyrene/divinyl benzene
Ligand:
Phenyl
Bead form:
Rigid, spherical porous, monodisperse
Particle size:
15 µm
Capacity:
At least 40 mg albumin/column
pH stability:
Long term
Short term
2–12
1–14
Temperature:
Regular use
Storage
+4 to +40 °C
+4 to +30 °C
Pressure over the column
Regular use
Do not exceed
0.25–2.5 MPa
4 MPa
Flow rate:
Regular use
Maximum
(water at room temperature)
0.5–2.5 ml/min
5.0 ml/min
First time use
Equilibration of the column before initial use or after long term storage
or changing buffers:
1.
8 ml elution buffer.
2.
8 ml starting buffer.
Starting buffer:
50 mM phosphate buffer, 1.5 M (NH4)2SO4, pH 7.0
Elution buffer:
50 mM phosphate buffer, pH 7.0
Flow rate:
1.0 ml/min
Note:
Before connecting the column, start the pump and remove
all air in the system, in particular in tubings and valves.
Try these conditions first
Before applying the sample, equilibrate the column.
Proceed according to the section “First time use”.
Starting buffer*:
50 mM phosphate buffer, 1.5 M (NH
4
)
2
SO
4
, pH 7.0
Elution buffer:
50 mM phosphate buffer, pH 7.0
Gradient:
0-100% B in 33 ml (20 CV)
Flow rate:
2.0 ml/min
*
Use a lower concentration of ammonium sulphate if the protein of interest
begins to precipitate at this concentration.
Read the back of this instruction for information
on optimising a separation.
Injection
valve
Monitor
Bed length 100 mm, i.d. 4.6 mm
5 µm
Buffer and solvent resistance
De-gas and filter all buffers through a 0.22 µm filter.
Instructions
SOURCE 15PHE PE 4.6/100
Daily use
Aqueous solutions pH 2–12
Urea, up to 8 M
Acetonitrile, up to 30% in aqueous buffers
Cleaning
Acetonitrile, up to 30%
Ethanol, up to 100%
Methanol, up to 100%
2-propanol, up to 100%
Hydrochloric acid, up to 1 M
Sodium hydroxide, up to 2 M
Acetic acid, up to 50%
Guanidine hydrochloride, up to 8 M
Anionic, cationic and non-ionic detergents
Avoid
Unfiltered solutions
Oxidising agents
Sample requirements/recommendations
Recommended
≤
40 mg protein/column
sample load:
Sample preparation:
Filtered through a 0.22 µm filter or
centrifuged at 10 000 g for 10 min
Temperature*:
Ambient
The sample should be dissolved in starting buffer.
*Hydrophobic interactions increases with increased
temperature. Results achieved at room temperature
may therefore not be reproduced in cold room, or
vice versa.
In Depth Information
Delivery/storage
The gel is delivered in 20% ethanol. If the column is to be stored for
more than two days after use, wash the column with at least 8 ml
distilled water and then equilibrate it with at least 8 ml 20% ethanol.
