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Multicolor detection is a powerful application of fluorescent Western blotting. Two or more proteins can be detected on one blot using antibodies labeled with different fluorophores (Figure 1). The ability to simultaneously assay multiple proteins...
The advent of digital imaging has transformed Western blot detection. Here we outline how Western blotting has changed and review qualities to look for in a digital imager. Chemiluminescence and fluorescence When most people think of imaging their...
Azure Biosystems is dedicated to designing instruments that can deliver industry-leading performance across a broad range of laboratory applications without overcomplicating the user experience. We have designed a suite of upgradeable cSeries inst...
Detecting in-gel fluorescence enables the analysis of protein gels immediately after electrophoresis without staining or Western blotting. Separation of proteins by electrophoresis is a cornerstone of protein analysis. After separation identificat...
Introduction Stain-Free gel technology enables detection of protein bands in gels and on transfer membranes without using colorimetric or fluorescent stains. Stain-Free gels (Bio-Rad) contain a trihalocompound within the gel matrix that produces a...
Chemiluminescent Westerns Chemiluminescent Western blotting is an indirect method for detecting proteins bound on a membrane. The method relies on an enzyme-substrate reaction that emits light which is traditionally detected on x-ray film. Chemilu...
Normalization is critical for obtaining accurate quantitative data from Western blots. Normalization accounts for unequal loading of samples across the lanes on a gel and for differences in transfer efficiency across a blot. Traditionally protein ...
Introduction Change is hard. It takes work to adapt to a new technology and to get used to something different. It’s especially hard if you have something that already seems to work. This is a big reason people still use film and darkrooms to proc...
Fluorescent Western blots are the gold standard for quantitative Westerns. They are ideal for detecting multiple proteins simultaneously (multi-plexing) allowing in-lane normalization and detection of your protein of interest and loading control a...
Chemiluminescent Westerns are popular assays for assessing protein expression. In this indirect detection method chemiluminescent substrates emit light when reacted with an antibody conjugated to an enzyme. The emitted light is captured and archiv...