![]() Here we provide data that define what should be considered, what avoided and what documented when quantifying Western blots. It constitutes a multistep method that allows the detection and/or quantification of a protein of interest and protein post-transcriptional modifications from different samples (Taylor et al., 2013). The Southern blotting technique was named after. It is a multi-step process, that begins with the electrophoresis of DNA, transfer of DNA fragments into nitrocellulose strip, and exposing those fragments to a DNA probe labeled with a radioactive or chemical tag. In the last decade, it has become apparent that assumptions underpinning these comparisons are often violated in studies reporting Western blot data in the literature. After the first detailed description of Western blot, this technique has been widely used in molecular labs for decades. Southern blotting is a molecular technique to find target DNA sequences in a sample. At present, the current lack of any definitions in densitometry opens the door to uncontrollable acquisition of any desired p-value. Densitometry data generated for Western blots are commonly used to compare protein abundance between samples. The Western blot protocol begins with the separation of larger molecules via electrophoresis. Western blotting or immunoblotting can pinpoint one or more specific protein antibodies from a sample via a standard protocol. Different common densitometry procedures applied to the identical Western blot revealed p-values of these correlations ranging from 0.000013 to 0.76 reflecting the necessity of a scientifically sound basis for densitometry of Western blots. The Western blot test detects proteins of the immune system and has been used since the late 1970s. The importance of such information, however, becomes evident from our correlations of plasma erythropoietin values of various mammals determined using RIA and Western blot densitometry. Southern blotting involves such important develop- ments as: the discovery of restriction enzymes the application of gel electrophoresis to the study of DNA the use of radioactive DNA probes to detect DNA sequences in a highly specific manner. A variety of techniques are available for the quantification of proteins, their degradation products and other impurities. Confirming previous experience, none of 100 randomly selected and systematically scanned most recent papers provided sufficient information on how Western blot results were translated into statistical values. Although Western blots are frequently quantified, densitometry is not documented and appears to be based merely on traditions and guesswork.
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