Biomedical Engineering Reference
In-Depth Information
rinsed with water. Soln. B is substituted by Soln. C and incubation
is done for several hours or overnight with gentle agitation. Then
the sheet is carefully rinsed with ddH
2
O and dried. Proteins are
visible as reddish bands.
References
Moeremans W, Daneels G, De Mey J (1985) Anal Biochem 145:315
2.4.5.3 Staining on PVDF Blotting Membranes with Dyes
Proteins blotted on PVDF membranes are stainable with Coomassie
Brilliant Blue R250, but a relative intense background remains,
which does not influence, for example, amino acid sequence anal-
ysis.
A
0.1% Coomassie Brilliant Blue R250, 40% Methanol (v/v), 1%
Solutions/Reagents
acetic acid (v/v) in ddH
2
O
B 80% methanol (v/v) in H
2
O
C 0.005% Sulforhodamin B (Xylylene Red B, Acid Red 52, C.I.
45100) (w/v), 30% methanol (v/v), 0.2% acetic acid (v/v) in
H
2
O
Methanol
Staining with Coomassie Brilliant Blue R250
ThemembraneisrinsedwithH
2
O after protein transfer (electro-
transfer or dot blot) and dipped to methanol for some seconds.
Immediately after this (avoid drying), the membrane is agitated
in Soln. A for maximum 2 min and destained by several changes
of Soln. B. When the membrane is dry, blue bands are visible on
a slight blue background. Membranes stained by this method are
not suitable for any immunochemical detection.
Staining with Sulforhodamin B
After blotting, the membrane is washed with ddH
2
Otwotimes
10 min to remove salts. The membrane is dried overnight or by
vacuum at RT. The dry membrane is incubated in Soln. C for 1 -
2 min, rinsed with ddH
2
O, and dried.
CPTS is also suitable for staining of proteins on PVDF mem-
branes (see Protocol 2.4.5.1).
References
Hancock K, Tsang VCW (1983) Anal Biochem 133:157
