shRNA gene silencing Human THP-1

Protein ladders are a set of standards known as molecular weight proteins that are utilized to identify the approximate size of a protein molecule run on a PAGE gel electrophoresis. The challenges in running the ladders are the choice of appropriate protein standard as it is used as visual evidence of protein migration, transfer efficiency, and positive control. Suitable protein markers can be selected on the basis of required properties and applications, i.e., fluorescent ladder, IEF, 2D SDS-PAGE ladder, natural ladder with an isoelectric point, and optimized ladders for Western Blot chemiluminescence detection. The key factors for running a distinct protein ladder are buffer conditions, charge/voltage at migration time, and the gel's concentration.

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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