Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 29)

Total Protein Blot Stains
Method Sensitivity Optimal Protein Load Advantages Disadvantages
SYPRO Ruby protein blot 2–8 ng ~0.2 µg/band Compatible with mass Multiple step protocol; Requires
stain spectrometry, Edman-based imaging instrument for
sequencing, and standard
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
AmidoBlack 10B 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 (HRP) 100 pg ~0.05 µg/band Color does not fade More expensive than 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 antigen May detect endogenous
(AP) enzyme activity
Amplified alkaline 10 pg ~0.005 µg/band High sensitivity Amplification requires
phosphatase 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
Immun-Star™ 1–3 pg ~0.005 µgband Intensifies signal Requires visualization on
chemiluminescent (hrp) output, very sensitive film or instrumentation
Immun-Star WesternC (HRP) 10 fg ~0.005 µgband long-lasting signal Requires visualzation on
short and multiple on film or instrumentation
exposures possible
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