The colour change of
redprot
®
solution from light pink to deep blue, which is induced by protein-dye interaction,
referes to a massive change in the spectrum near 600 nm wave length.
Spectral changes of the redprot® staining fluid
1. Staining of complex mitures of proteins
Change in absorption of redprot®
caused by interaction with complex protein mixture
in extracts of different pollen
2. Staining of proteins with
redprot®
in cells : spectra of noumerous pollen measured in situ
Figure 1: Artemisia officinalis stained with
redprot®
Figure 2: Betula vericosa stained with
redprot®
Figure 3: Lolium spec. stained with
redprot®
Figure 4: Phleum spec. stained with
redprot®
The redprot® solution contains a dye-metal ion complex:
- Pyrogallol red (PR) + Molybdate (Mo) = dye-metal complex
- interaction with Protein = PR-Mo-Protein complex
- protein-selective stain: electrostatic interrelations with more than one residue
- primarily via basic amino acids - minor contribution of aliphatic residues
- protein interaction induces spectral changes around 600 nm
Figure: Spectral effect on redprot® dilution by aqua dest.
Figure: There is no effect of buffer salts visible in the spectrum at 600 nm wave length
Biochemical application of redprot® compared to Coomassie Brilliant Blue
| Application | redprot® |
Coomassie Blue |
| Protein Assay Detection range: |
10 µg to 16mg/ml | 5 µg to 2 mg/ml |
| Membrane Staining Detection limit background destaining |
10 ng per spot on NC and PVDF free poor interference with agents mild with water |
100 ng per band on PVDF Permanent high destaining longlasting with acidic alcohol solutions |
| Gel Staining | Possible, less sensitive: 1µg/lane | Highly sensitive 30 ng |
| Protein precipitation | quantitaive and non-denaturating: min. concentration1µg/ml |
Not possible |