InstantBlue is a ready-to-use, proprietary Coomassie® stain that is specially
formulated for ultra-fast, sensitive and safe detection of your proteins. Protein
gels can be stained in minutes without the need to wash, fix or destain.
Only proteins are stained resulting in well defined blue bands on a highly
transparent background.

The reduction of background interference results in a
better signal to noise ratio and may also have a positive impact on the overall
resolution and sensitivity.
The InstantBlue formulation is non-toxic and does not contain any methanol.
Proteins stained using the InstantBlue stain are also compatible with mass
spectrometry (MS) analysis.

• Contents: 1L reagent, containing Coomassie dye, ethanol, phosphoric acid and solubilizing
  agents in water. (Caution: Phosphoric acid is a corrosive liquid.)
   
• Storage
  Upon receipt store at + 4°C. Discard any reagents that show discoloration or
  evidence of microbial contamination. Be sure to keep the bottle capped when not
  in use.
   
• Before Use :
  Mix the InstantBlue solution immediately before use by gently inverting the bottle a few
  times (do not shake the bottle to mix the solution).
   
• IMPORTANT  1) Multiple washes prior to staining with InstantBlue are NOT
  required or recommended.
  2) An alcohol/acetic acid fixing step prior to staining with
  InstantBlue is NOT required or recommended.
  3) A destaining step post staining is NOT required or
  recommended with InstantBlue.
  Standard Protocol :
  1) After electrophoresis remove the gel from the tank and transfer directly into the
  InstantBlue staining solution. Be sure that the gel moves freely in stain to
  facilitate diffusion. Typically ~20 ml is needed to cover the gel.
  2) Coloured protein bands will start to develop immediately and a suitable intensity
  is typically achieved after 15 minutes incubation at room temperature with
  gentle shaking.
  3) Photograph your gel when the required intensity has been achieved. Gels can be
  kept in staining solution, but ensure that the gel remains covered with liquid.
  Close container to reduce evaporation of InstantBlue. Alternatively the gel can
  be stored in ultrapure water after staining for 1 hour in InstantBlue.
  4) Once used, the staining solution should be discarded and cannot be reused.
  InstantBlue is provided as ready-to-use solution and should not be diluted.

Protocol for Gel Drying :
1) Ensure that the gel has been staining for at least 1 hour.
Although protein bands will be visible after a few minutes of incubation in stain,
the staining process is typically fully completed after 1h incubation. Depending
on the type of gel you are using longer incubation may be necessary. Further
processing of the gel prior to completion of the staining process may result in
protein destaining and reduced sensitivity. If this occurs simply restain the gel
by incubating overnight in InstantBlue.
2) Submerse the gel in approximately 100 ml ultrapure water at ~70°C (heat for 30s
to 60s in a microwave oven). Incubate for at least 1 hour while gently rocking.
Optionally adsorbent paper or paper towel can be added. Gels can be incubated
overnight in water.
3) Incubate the gel in a ‘gel drying solution’ (e.g. 4% glycerol, 20% ethanol in
water) for 2 minutes. Incubation of any Coomassie®-stained gel in an alcohol
solution will eventually result in destaining of the bands so avoid incubation for
longer than 5 minutes.
4) The gel is now ready for drying between wetted cellophane membranes.
Protocol for Destaining Protein Bands for MS analysis :
1) Excise the protein band of interest and transfer to a clean Eppendorf tube.
2) Add 1 ml of 30% ethanol or 30% acetone or 30% acetic acid
(Note: Acetic acid may result in acetylation of the N-terminus)
3) Incubate for 20min (incubate at 60°C – 70°C to increase the rate of destaining)
4) Decant supernatant and repeat step 2&3 at least 3 times or until gel is clear
For more detailed protocols please contact your MS facility