Quantify with Confidence. Stain Once, Blot Accurately.
Description
AdvanStain Total is a highly sensitive fluorescent protein stain for PVDF and Nitrocellulose membranes that delivers clear, quantitative visualization of your protein bands. To achieve truly precise Western blot data, total protein normalization is no longer optional - it's the standard preferred by top-tier journals for maximizing experimental transparency and accuracy.
The kits provide a convenient, robust solution for this essential step. Compatible with any digital imaging system equipped with a Cy3-compatible filter, AdvanStain Total ensures your normalization data is sensitive, reliable, and perfectly suited for your most critical quantitative analysis. Elevate your blots from qualitative images to peer-review ready data.
- 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
- for 15 miniblots
AdvanStain total spectrum
AdvanStain Total can be imaged with a CCD or laser based imaging system using green-light excitation.
Broad linear range
AdvanStain Total has a broad linear range. Untreated Hela lysate was serially diluted 1:2 prior to separation by SDS-PAGE. After electrophoresis the proteins were transferred to a PVDF membrane for total protein staining (a). The image was quantified, and staining intensities analyzed by linear regression (b).
Fluorescent Western Blotting
AdvanStain Total is compatible with IR fluorescent Western blotting, without destaining. 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
AdvanStain Total is compatible with chemiluminescent Western blotting, without destaining. 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
AdvanStain Total may be imaged wet or dry without increasing general background. 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).