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Western blots provide the ability to detect specific protein expression levels from cells or tissues in a convenient format for rapid evaluation that can also be quantitative and offer unparalleled sensitivity. With a variety of detection techniques (chemiluminescence, fluorescence, or colorimetric) to choose from, you can select a technology to match your experimental requirements and the instruments you have available. 

Which western blot detection technology that is right for you?

 
  Chromogenic Detection Kits Chemiluminescent HRP & AP Detection Kits Alexa Fluor Conjugates Western Dot Fluorescent Conjugates
Detection Type Chromogenic Chemiluminescence Fluorescence Long-wavelength Fluorescence
Sensitivity + +++++ ++ +++
Signal Duration Months Hours Months Years
Multiplexing No No Yes Yes
Means of Detection By eye; no instrument required X-ray film, ccd imagers Fluorescent digital imager, NIR Imaging system Fluorescent digital imager, UV -transilluminator, UV-gel documentation systems, direct
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Chromogenic Western Blot Detection

Chromogenic or precipitating substrates have been used widely for many years and offer the simplest and most cost-effective method of detection. When these substrates come in contact with the appropriate enzyme, they are converted to insoluble, colored products that precipitate onto the membrane. The resulting colored band or spot requires no special equipment for processing or visualizing. Chromogenic blotting substrates are available in a variety specifications and formats. The appropriate substrate choice depends on the enzyme label, desired sensitivity and form of signal or method of detection needed.

Chemiluminescent Western Blot Detection

Chemiluminescent substrates are popular because they offer several advantages over other detection methods. These advantages have allowed chemiluminescence to become the detection method of choice in most protein laboratories. Using chemiluminescence allows multiple exposures to be made in order to obtain the best image. The detection reagents can be removed and the entire blot reprobed to visualize another protein or to optimize detection of the first protein. A large linear response range allows detection and quantitation for a large range of protein concentrations. Most importantly, chemiluminescence yields the greatest sensitivity of any available detection method.

Fluorescent Western Blot Detection

Fluorescent reagents are growing in popularity for Western blotting because they offer increased time savings over chemiluminescent detection and reduced chemical waste compared to both chemiluminescent or chromogenic detection systems. While the detection limits are still not as low as chemiluminescent detection, fluorescent detection has the unique advantage of allowing multiple targets to be assayed for on the same blot at the same time without the need to strip and reprobed.

SuperSignal Western Blot Signal Enhancer

Offers increased sensitivity, specificity, and clarity in your Western blots.

The Thermo Scientific SuperSignal Western Blot Enhancer contains a membrane treatment reagent and a primary antibody diluent that increase both signal intensity and sensitivity 3- to 10-fold compared to detection performed without it.