How to choose the appropriate western Blot imaging method The most appropriate Western blot method depends on what information you expect, and the following will help you choose the best method of experimentation. Chemiluminescence method Chemiluminescence western blot is a relatively easy and highly sensitive detection method with sensitivity up to fg. Proteins with significant molecular weight differences can be easily separated by electrophoresis, and it is very convenient to use "chemiluminescence" detection. Advantage: Disadvantages: high sensitivity Easy, familiar with the experimental protocol Compatible film and digital imaging Semiquantitative analysis Enzymatic reaction Only single signal acquisition is possible, and sample loading requires stripping and secondary incubation Protein gel electrophoresis: Detection of a single protein Determine if the protein is present Detection of antibody response Protein purity analysis Detection of low abundance proteins Chemiluminescence, single signal, single protein Fluorescence method Fluorescent Western blot was detected by fluorescent dye-labeled secondary antibody, and the target protein concentration on the blotted membrane was linear with the fluorescence signal intensity, realizing reliable and quantitative analysis. Fluorescent dyes have different emission spectra, so multiple detections can be performed on one blot. It is especially useful when analyzing the post-translational modified protein without the need for stripping and secondary incubation. Fluorescent western blots use infrared or visible fluorescent dyes. Advantage: Disadvantages: Multiple detection High quantitative accuracy Fluorescent near-infrared blotting membrane for storage and repeatable imaging Sensitivity is slightly lower than chemiluminescence method Film autofluorescence leads to increased background noise Visible fluorescence detection advantages When three proteins need to be detected simultaneously, it is apparent that fluorescence is the best choice. Visible fluorescence is very flexible and there are a variety of visible fluorescent dyes to choose from. If you are using other visible fluorescent dyes (the emission spectra can be distinguished) for other experiments, it is easy to apply these dyes to Western blot experiments. Fluorescence detection method can be used to: Simultaneous detection of multiple proteins Post-translational modification protein Quality control of the same imprinted membrane Normalization of multiple lanes Precise quantification Western experiment Multiple fluorescence, multiple proteins Shanghai Jiecheng Biotechnology Co., Ltd. http://
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