1 ph dependence – Bio-Rad Affi-Gel 15 Gel User Manual
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Section 3
General Coupling Conditions
3.1 pH Dependence
A major advantage of Affi-Gel 10 and 15 supports is
the mild conditions which will permit coupling. This is
particularly advantageous in applications which involve
sensitive enzymes or other proteins that irreversibly lose
biological activity when exposed to conditions outside
their physiological range. Coupling can be achieved with
Affi-Gel 10 and 15 supports between pH 3.0 to 10.0.
In order to maintain pH control, a minimum buffer
strength of 10 millimolar is recommended. Suitable
buffers include MES, MOPS, HEPES, POPSO, acetate,
and bicarbonate. Do not use buffers such as Tris or
glycine. They contain primary amino groups which will
couple to the gel, as will any primary amine-containing
compound which contaminates the ligand preparation.
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Product Description for Affi-Gel 10 and 15 gels
Matrix
Bio-Gel A-5m agarose gel
Exclusion limit (M
r
)
5,000,000
Bead size
75-300 µm (50-200 mesh)
Spacer arm
Affi-Gel 10
Affi-Gel 15
Shipping medium
100% isopropanol
Capacity
Chemical capacity
15 µmoles/ml of gel
Protein capacity
35 mg/ml
Stability of unreacted support
Temperature
-70 to 0 °C
pH range
3-10
Organic solvents
stable in alcohols, DMSO, dioxane, formamide
Storage
-20 °C 1 year
-70 °C 1.5 years
OCH
2
CONH(CH
2
)
3
N(CH
2
)
3
NHCO(CH
2
)
2
COON
O
O
CH
3
H
+
OCH
2
CONH(CH
2
)
2
NHCO(CH
2
)
2
COON
O
O
4
LIT156B 6/17/98 10:41 AM Page 4