Reversible staining of proteins on PVDF membranes

Summary

Reversible staining of proteins on PVDF membranes can be used for: confirming the transfer of proteins to PVDF membranes during Western hybridization.

Operation method

Reversible staining of proteins on PVDF membranes

Principle

Lichtenstein Red is negatively charged and can bind to positively charged amino acid residues, as well as to non-polar regions of proteins, resulting in a reddish band.

Materials and Instruments

Protein
AmidoBlack isopropanol aceticacid Kaomas Brilliant Blue methanol Ponceau S in TCA EtOH HAC Na2S2O3 AgO3 Na2CO3 MeOH AgNO3 formaldehyde
PVDF membrane

Move

1. Amido Black staining

Dyeing solution: 0.5% Amido Black (w/v), 25% isopropa nol (v/v) and 10% acetic acid.

Staining: Place the PVDF membrane in the staining solution for several hours, and decolorize with _ddH2O.

2. Kaomas Brilliant Blue Staining

Staining solution: 0.1% Coomassie Brillia nt Blue R-250 (w/v) and 50% met hanol (v/v)

Decolorizing solution: 40% met hanol (v/v) with 10% acetic acid (v/v)

Staining: Place the PVDF membrane in the staining solution for 15 min. and decolorize with decolorizing solution.

3. Lichun red staining Ponceau S

Dyeing solution: 0.2% w/v Ponceau S in TCA (3% v/v)

Staining: Place the PVDF membrane in the dye solution for 5min.

Decolorization: _ddH2O decolorization

Caveat

Lichtenstein Red Stain is not suitable for the detection of proteins on nylon membranes.

Common Problems

The staining sensitivity of Lichtenstein Red is not as good as that of the Caulmers Brilliant Blue staining solution.

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