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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
: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 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 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 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 cleartimes orFor more detailed
protocols please contact your MS facility
Protein Detection & Quantitation:
Determination of Protein Concentration
NVoy Technology in Protein Circular Dichroism (CD)
NVoy Technology Improves Protein Recovery on PD10 Desalting Columns
NVoy Technology Facilitates Protein Mass Spectrometry
NV10 Improves Recovery From Protein Concentration
The Application of NVoy Technology to Protein Stabilisation
Stabil-P.A.C. NV10 Improves Protein Recovery after Dialysis
Stabil-P.A.C. NV10 Stabilises Fusion Proteins after Tag Cleavage
To view our other products let us
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Gentaur /
Clonagen /
Genoprice / Bioxys /
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