A5101 Datasheet, PDF (1/1 Pages) List of Unclassifed Manufacturers – IMMOBILIZED β -GALACTOSIDASE F7M

PRODUCT INFORMATION SHEET
IMMOBILIZED β -GALACTOSIDASE F7M
# A5101
β-galactosidase from Escherichia coli.
β-galactosidase catalyzes the hydrolysis of β-D-galactoside to galactose and alcohol.
F7m: 1.0 mg β-galactosidase immobilized on polyvinyl ,
780 units immobilized per CR-column.
100 µl of a 5% lactose solution is digested to over 80% in 60 minutes
Nr.14 Storage buffer:
Nr.64 Reaction buffer:
Nr.63 Washing buffer:
50 mM Tris/HCl, pH 7.8
add DTT to the storagebuffer, final concentration: 1 mM DTT
50 mM phosphate, 1 mM MgCl₂, pH 7.8 add β-mercaptoethanol,
to the reactionbuffer, final concentration: 10 mM
50 mM phosphate, 1 M NaCl, pH 7.8
Protocol
For more details see MoBiTec-CRC-Handbook.
1. Dilute delivered buffers (at least 2 ml each) with sterile doubly distilled water.
For 1 application you need
0.25 ml 10x reaction buffer and 2.25 ml doubly distilled water+
10 mM β-mercaptoethanol
0.4 ml 5x washing buffer and 1.6 ml doubly distilled water
0.2 ml 10x storage buffer and 1.8 ml doubly distilled water+
DTT (1 mM )
The substrate should be in reaction buffer
2. Equilibrate the CR-column with 2 ml reaction buffer.
Fill 2 ml reaction buffer into a syringe, let the reaction buffer run through the column by
gravity to the upper filter. In case the buffer runs very slowly, apply pressure by a syringe.
3. Load substrate solution in reaction buffer.
Small volumes (< 80 µl): spin the CR-column 5 seconds in a benchtop centrifuge (2000 rpm
are sufficient). Let the substrate solution enter the matrix material.
Larger volumes:
Let the substrate solution run through the column.
Flow-rate:
up to 80 µl/minute
Keep the substrate in the column for about 1 minute at room temperature. Higher turn-over
is obtained when the substrate is applied to the column again or incubated for longer times.
4. Elute the product solution.
Small volumes (< 80 µl): Elute the product with 500 µl reaction buffer.
Larger volumes:
Let the substrate run through the column and elute the residual
product solution with 500 µl reaction buffer.
It does not harm the columns if they run dry.
5. Wash the column with 2 ml washing buffer.
6. Equilibrate the column with 2 ml storage buffer.
Store the column at 4°C.
Never freeze a CR-column!
MoBiTec # A5101 02/10/2000
For research use only. Not for use in diagnostic or therapeutic procedures.
MoBiTec GmbH, Lotzestraße 22a, D-37083 Göttingen, Tel.: ++49 551-70 722- 0, Fax: ++49 551-70 722-22
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