Instant-Bands Prestaining Protein Sample Treatment Buffer for SDS-PAGE
Instant-Bands sample treatment buffer (sample loading buffer) pre-stains a protein sample for SDS-PAGE. Protein samples are stained fluorescently during sample treatment prior to electrophoresis. Protein bands in an SDS-gel and on a membrane after transfer can be visualized and analyzed by an UV or LED transilluminator or by a digital gel image system. Instant-Bands are more sensitive than silver staining.
Instant-Bands have been formulated to enhance the sensitivity, stability and to eliminate background. Instant-Bands stain all proteins. It doesn’t change protein migration rates and electrophoresis patterns. During electrophoresis, free dye molecules migrate at the same rate as the tracking dye bromophenol blue and migrate to the bottom of the gel at the end of a run, leaving a clean background. Instant-Bands ideally suit to an SDS-PAGE experiments for tracking protein expression and purification and Western blot.
• No more gel stain/de-stain
• No special instruments or reagents required
• No extra experimental steps
• No need to retrieve gels from gel cassettes
• Bands retain on membranes after transfer
• Compatible with Western blot
• Stain the selected lanes, not the entire gel
• Apply to any gels, hand-made or precast
1. Mix Instant-Band Sample Treatment Buffer with a protein sample in 1:2 ratio (volume).
For example, mix 3 μl Instant-Bands with 6 μl protein sample.
2. Heat the sample at 90-100 ºC for 10 minutes.
For whole cell or tissue samples, increase the heating time to 15 minutes. Make sure your heating block is >90oC to let the sample be sufficiently heated.
3. The sample is ready for gel electrophoresis.
If the molecular weight marker is needed, load 3 μl ready-to-use pretreated fluorescent
molecular weight makers to a sample well.
4. After electrophoresis, place the gel on a transilluminator to view protein bands and take pictures.
The transilluminator can be an UV, a LED blue or a digital gel image system. The gel does not need to be removed from the gel cassette if light source is in visible wavelength range.
5. (Optional) The gel can still be stained by Coomassie Blue if desired: Stain the gel according to the standard Coomassie Blue protocol