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Choose an appropriate protein gel (SDS-PAGE) stain

Once protein bands have been separated by electrophoresis, they can be visualized using different methods of in-gel detection. Each staining method has particular advantages and disadvantages, including overall convenience, detection limit and compatibility with downstream applications and detection instruments.

The following charts provide a general guide to selecting a protein stain for a given application based on detection method, sensitivity and compatibility with mass spectrometry.

Total Protein Stains

Thermo Scientific Gel Stains for detection of total protein.

Some products require additional reagents to be supplied by user. The number of steps includes multiple gel washes, solution changes, destaining steps, etc. Times include gel wash steps and all steps required for best sensitivity in a mini-gel. When less sensitivity is required, shorter staining and destaining times may be used with some protocols.

Product Components
(steps)
Time
required
Detection
method
Limit of
detection
Mass spec.
compatible
Krypton Fluorescent Protein Stain 1 (7) 2 hr 40 min fluorescent 0.25 ng Yes
SYPRO® Ruby 1 (3) 90 min microwave
18 hr standard
fluorescent 0.25 ng Yes
SYPRO® Orange 1 (5) ~1 hr fluorescent 4-8 ng  
SYPRO® Red 1 (5) ~1 hr fluorescent 4-8 ng  
SYPRO® Tangerine 1 (4) ~1 hr fluorescent 4 ng Yes
Coomassie™ Fluor Orange 1 (4) ~1 hr fluorescent 8 ng  
GelCode Blue Safe Protein Stain 2 (5) 2 hr 15 min colorimetric 9 ng Yes
GelCode Blue Stain Reagent 1 (5) 2 hr 15 min colorimetric 8 ng Yes
Imperial Protein Stain 1 (5) 2 hr 15 min colorimetric 3 ng Yes
Simply Blue™ Safe Stain 1 (5) 12 min microwave
3 hr standard
colorimetric >7 ng Yes
PageBlue Protein Stain 1 (5) 30 min microwave
90 min standard
colorimetric 5 ng Yes
Pierce Silver Stain for Mass Spec 6 (17) 1 hr 13 min colorimetric 0.25 ng Yes
Pierce Silver Stain Kit 4 (15) 2 hr 25 min colorimetric 0.25 ng Yes
Pierce Color Silver Stain Kit 4 (9) 4 hr 5 min colorimetric 0.1 ng Yes
Pierce Zinc Reversible Stain Kit 3 (4) 14 min colorimetric 0.25 ng Yes
SilverXpress® Silver Staining Kit 5 (13) 2 hr colorimetric 0.86 ng Yes

Functional Group Protein Stains

Thermo Scientific Gel Stains for detection of specific classes of proteins. Some products require additional reagents to be supplied by user. The number of steps includes multiple gel washes, solution changes, destaining steps, etc. Times include gel wash steps and all steps required for best sensitivity in a mini-gel. When less sensitivity is required, shorter staining and destaining times may be used with some protocols.

Product Components
(steps)
Time
required
Detection
method
Limit of
detection
Mass spec.
compatible
InVision™ His-tag In-gel Stain 2 (4) <3 hrs UV ~0.5 picomole of a 6X His-tagged fusion protein not tested
Lumio™ Green Detection Kit 3 (8) Immediate UV 1 pmole of a Lumio™ fusion protein not tested
6xHis Protein Tag Stain Kit 4 (9) 2 hr 20 min fluorescent 200 ng of 35 kDa
His-tagged protein
not tested
Pro-Q® Emerald 488 Glycoprotein Gel and Blot Stain Kit 2 (13) ~6 hrs fluorescent 4 ng glycoprotein / band Yes
Pro-Q® Emerald 300 Glycoprotein Gel and Blot Stain Kit 2 (10) ~5 hrs fluorescent 0.5 ng glycoprotein / band Yes
Pro-Q® Diamond Phosphoprotein Gel Stain 1 (5) ~4-5 hrs fluorescent 1–16 ng phosphoprotein / band not tested
Multiplexed Proteomics™ Phosphoprotein Gel Stain Kit varies ~4-5 hrs fluorescent 1–16 ng phosphoprotein / band  
Pierce Glycoprotein Stain 5 (11) 2 hr 40 min colorimetric 160 ng not tested
Phosphoprotein Staining Kit 9 (12) 3 hr 45 min
+ overnight
destain
colorimetric 80 ng phosvitin,
160 ng beta-casein
not tested