The Protein Man's Blog | A Discussion of Protein Research

The Protein Man

The Protein Man

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How to Prepare Samples for Western Blot Analysis

Posted by The Protein Man on Jan 21, 2020 2:30:00 PM

As a common technique in molecular and cell biology, Western blot analysis allows you to detect specific proteins in a tissue homogenate or extract. However, if you want to ensure the success of your Western blot experiment, you need to learn how to properly prepare samples.

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Topics: Western Blotting

What’s the Difference Between Tween 20 and Tween 80?

Posted by The Protein Man on Jan 14, 2020 2:30:00 PM

As a non-ionic viscous liquid, tweens are a class of emulsifiers used in pharmaceuticals and food preparation.

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Topics: Detergents

The Role of LDH in Cellular Cytotoxicity

Posted by The Protein Man on Jan 7, 2020 2:30:00 PM

Analyzing the effects of necrosis and apoptosis is a vital component of biological research. Although there are numerous assays to detect apoptosis, there are significantly fewer assays available to measure necrosis. The release of LDH is a beneficial method for detecting necrosis, especially when combined with cellular cytotoxicity.

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Topics: Apoptosis Assays, Cytotoxicity Assays

Bradford Assay for Protein Concentration

Posted by The Protein Man on Dec 3, 2019 2:30:00 PM

The Bradford assay is a fast and efficient colorimetric assay to determine the protein concentration in a wide range of solutions and is widely used for protein content of cell fractions and protein concentrations for gel electrophoresis. It is often done before an SDS-PAGE analysis. Using the reagent Coomassie Brilliant Blue G-250, the solution undergoes a visible change in color when the dye binds to proteins via van de Waals and electrostatic interactions, causing a shift from a dark red to a deep blue. This is a result of stabilization moving the absorbance maximum of the dye from 470 to 595 nm. Tested samples typically take less than five minutes to reach the indicative measures of 595 nm absorbance concentration, making the Bradford assay both reliable and quick.

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Topics: Protein Estimation

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