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ProStain™ simplifies the determination of cell extract protein concentrations by providing highly sensitive detection reagents that are easy to use. The dye used in ProStain is resistant to many commonly found contaminants in cell lysis buffers such as detergent and salt, which limit the usefulness of other protein quantification systems. And, the spectral properties of ProStain change greatly after it is bound, effectively eliminating any background from unbound dye (see Properties tab). These characteristics make the ProStain Kit a significant improvement over other detection systems.
The background of ProStain is extremely low because its absorption max shifts by over 100 nm after the dye is bound, and because bound dye has a 50-fold greater quantum yield than free dye (Figure 1, Table 1).
Figure 1: Absorption and emission spectra of free vs. bound dye.
Normalized absorption and emission spectra of free (solid, purple lines) and conjugated dye (dotted, copper lines) in phosphate buffer of pH 7.2. As free dye and bound dye absorb at different wavelengths (612 nm vs. 503 nm) and the quantum yield of bound dye is 50-fold greater than that of free dye, background from free dye is effectively eliminated.
| Dye State | Absorption | Emission | ε L/(mol-cm) | Quantum Yield |
|---|---|---|---|---|
| Free dye | 612 nm | 665 nm | 60,000 | < 1% |
| Conjugated dye | 503 nm | 600 nm | 24,000 | 50% |
