is a ready-to-use, proprietary Coomassie® stain that is specially
formulated for ultra-fast, sensitive and safe detection of your
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
The reduction of background interference
results in a
better signal to noise ratio and may also have a positive impact on the
resolution and sensitivity.
The InstantBlue formulation is non-toxic and does not contain any
Proteins stained using the InstantBlue stain are also compatible with
spectrometry (MS) analysis.
- Contents: 1L reagent, containing Coomassie dye, ethanol,
phosphoric acid and solubilizing
agents in water. (Caution: Phosphoric acid is a corrosive liquid.)
Upon receipt store at + 4°C. Discard any reagents that show
evidence of microbial contamination. Be sure to keep the bottle
capped when not
- 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
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
facilitate diffusion. Typically ~20 ml is needed to cover the gel.
2) Coloured protein bands will start to develop immediately and a
is typically achieved after 15 minutes incubation at room
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
Close container to reduce evaporation of InstantBlue. Alternatively
the gel can
be stored in ultrapure water after staining for 1 hour in
4) Once used, the staining solution should be discarded and cannot
InstantBlue is provided as ready-to-use solution and should not be
Protocol for Gel Drying :0080">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
the staining process is typically fully completed after 1h incubation.
on the type of gel you are using longer incubation may be necessary.
processing of the gel prior to completion of the staining prprotein 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
Optionally adsorbent paper or paper towel can be added. Gels can be
overnight in water.
3) Incubate the gel in a ‘gel drying solution’ (e.g. 4% glycerol, 20%
water) for 2 minutes. Incubation of any Coomassie®-stained gel in an
solution will eventually result in destaining of the bands so avoid
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
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
4) Decant supernatant and repeat step 2&3 at least 3 times or until gel
is cleartimes orFor more detailed protocols please contact your MS facility
send mail to firstname.lastname@example.org with questions or comments about this web site.
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