Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting
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چکیده
منابع مشابه
Immunofluorescent protein detection in Western blotting
This report describes the detailed procedures for Western blot analysis using fluorescent antibodies. After electrophoresis and subsequent electroblotting, the fluorescent-labeled antibodies were visible upon ultraviolet illumination of the polyvinylidene fluoride (PVDF) membranes and could then be photographed to give an accurate record of the blots. Fluorescent labeling allows for photographi...
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Reference proteins (RP) or the total protein (TP) loaded is used to correct for uneven loading and/or transfer in Western blotting. However, the signal sensitivity and the influence of physiological conditions may question the normalization methods. Therefore, three widely used reference proteins [β-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and α-tubulin], as well as TP loaded me...
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immunodetection is performed. Use of the Criterion Stain Free system eliminates the need for the timeand reagentconsuming traditional methods such as Ponceau S staining used to asses protein transfer prior to detection in western blotting applications. We also demonstrate that, while binding of some monoclonal antibodies may be affected by the Criterion Stain Free method, we have not observed a...
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ژورنال
عنوان ژورنال: Analytical Biochemistry
سال: 2014
ISSN: 0003-2697
DOI: 10.1016/j.ab.2014.08.027