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AdvanStain - Total Fluorescent Protein Staining Kits
AdvanStain Total is a fluorescent protein stain for PVDF and Nitrocellulose membranes that allows for sensitive and quantitative visualization of protein bands. For precise quantitative Western blot data, it is critical to perform normalization to account for sample inconsistencies, sample loading errors and uneven transfer. In an effort to increase transparency, total protein normalization is quickly becoming the standard for Western blot normalization as it is now the preferred method for top tier journals. The AdvanStain Total Fluorescent Protein Staining Kits provide a convenient method for total protein normalization that is compatible with digital imaging systems equipped with a Cy3-compatible filter.
Rapid fluorescent stain for total protein normalization
- Convenient – simple protocol that takes less than 20 minutes, no de-staining required
- Versatile – compatible with chemiluminescent and IR700/IR800 fluorescent Western blotting
- Accurate – linear across a broad range from 1-50μg of lysate, ideal for total protein normalization
Fluorescent Western blotting
Untreated and IFNα treated HeLa lysate was loaded in quadruplicate then transferred to a PVDF membrane using a wet transfer. After transfer, the membrane was stained with AdvanStain Total and imaged using a Cy3 filter (a). After staining, the blot was blocked and probed for GAPDH and phosphorylated STAT3. SpectraDye-IR700 and IR800 secondary antibodies were used for detection (b).
Chemiluminescent Western blotting
Untreated and IFNα treated HeLa lysate was loaded in quadruplicate then transferred to a PVDF membrane using a wet transfer. After transfer, the membrane was stained and imaged using a Cy3 filter (a). After staining the blot was blocked and probed for Actin and phosphorylated STAT3. The antibodies were detected with a goat anti-rabbit HRP conjugate and WesternBright-ECL HRP Substrate (b).
Maximum flexibility
Dilutions of untreated HeLa lysate were electrophoresed on an SDS-PAGE gel then transferred to a PVDF membrane using a wet transfer. After transfer, the membrane was partitioned and stained. AdvanStain Total was imaged in the Cy3 channel wet and dry (a), the competitor stain was imaged in the Cy5 channel wet and dry (b).