GE Amersham ECl Gel Box, Руководство пользователя

Страница: 60 / 72

Remedies Possible causes Problems
•
Select a gel that separates in
the desired molecular weight
range
•
Reduce loaded protein
amount
•
For proteins of similar
molecular weight, a 2-D
separation may be required
Incorrect gel selection, sample
overloading Bands difficult to
distinguish
•
Reduce salt by ultrafiltration
•
Optimize the amount of
protein applied to the gel
•
Excess salt in the sample
•
Too much protein applied to
the gel
Sample spreading
across the gel
•
Remove the carbohydrate by
enzymatic or chemical
means
Sample contains
appreciable
carbohydrate
•
Use a gradient gel
•
Try addition of a non-ionic
detergent
Sample contains
lipoproteins
•
Start with cold buffer (<15°C)
Excessive heating Protein
denaturation and
band inversion
•
Wash the gels extensively (3
x 10 minutes) with ultrapure
water and use 30% methanol
to destain the gel
•
Use 10% TCA to fix the
proteins in the gel. Add extra
reducing agent or change
the reducing agent.
•
SDS still present in the gel
•
Insufficient reduction of
samples
Diffuse protein
zones in the gel
after staining
60 Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC
7 Troubleshooting