Bio-Rad Criterion™ XT Tris-Acetate Precast Gels - Manual (Page 16)

Total Protein Blot Stain
Method
Sensitivity Optimal Protein Load Advantages Disadvantages
SYPRO Ruby protein 2–8 ng ~0.2 µg/band Compatible with mass Multiple-step protocol;
blot stain spectrometry, Edman-based Requires imaging
sequencing, and standard instrument for
immunological procedures maximum sensitivity
Colloidal gold stain 1 ng ~0.1 µg/band Sensitive, one step Not compatible with
nylon membranes
Enhanced colloidal 10–100 pg ~0.1 µg/band Increases sensitivity of Multiple steps
gold detection kit colloidal gold stain
Amido Black 100–1,000 ng ~5 µg/band Standard membrane Low sensitivity
stain, economical
Immunoblot Detection
Method
Sensitivity Optimal Protein Load Advantages Disadvantages
4CN colorimetric (HRP)* 500 pg ~0.25 µg/band Fast detection Results may fade
DAB colorimetric (HRP) 500 pg ~0.25 µg/band Fast detection Contains toxic
chemicals
Opti-4CN colorimetric 100 pg ~0.05 µg/band Color does not fade More expensive than
(HRP) 4CN
Amplified Opti-4CN ™ 10 pg ~0.005 µg/band High sensitivity, low Amplification requires
colorimetric (HRP) background additional steps
BCIP/NBT colorimetric 100 pg ~0.05 µg/band Sensitive; multiple antigens May detect endogenous
(AP) enzyme activity
Amplified AP* 10 pg ~0.005 µg/band High sensitivity Amplification requires
additional steps
Immun-Star ™ 10 pg ~0.005 µg/band Long-lasting signal, Requires visualization
chemiluminescent (AP) short and multiple on film or instrumentation
exposures possible
*(HRP) horseradish peroxidase; (AP) alkaline phosphatase
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