A Ponceau S Staining-Based Dot Blot Assay for Rapid Protein Quantification of Biological Samples
Dario Lucas Helbing, Leopold Böhm, Nova Oraha, Leonie Karoline Stabenow, Yan Cui
Abstract
Despite the availability of a wide range of commercial kits, protein quantification is often unreliable, especially for tissue-derived samples, leading to uneven loading in subsequent experiments. Here we show that the widely used Bicinchoninic Acid (BCA) assay tends to underestimate protein concentrations of tissue samples. We present a Ponceau S staining-based dot-blot assay as an alternative for protein quantification. This method is simple, rapid, more reliable than the BCA assay, compatible with biological samples lysed in RIPA or 2x SDS gel-loading buffer, and also inexpensive.
Topics & Concepts
Bicinchoninic acid assayChemistryLysisBradford protein assayProtein detectionMolecular biologyChromatographyWestern blotDot blotQuantitative analysis (chemistry)BlotLysis bufferBiochemistryIn vitroProtein expressionAdvanced Proteomics Techniques and ApplicationsMolecular Biology Techniques and ApplicationsAdvanced Biosensing Techniques and Applications