Cell lysis is a critical process for the study of intracellular proteins and nucleic acids. Cell lysis technique effectiveness can be measured by lysate yield and quality. Unfortunately, there are multiple points before and during the technique where the sample can be ruined, causing a waste of time, materials, and money. Here we have compiled specific ways to make the cell lysis process more efficient, so that you can achieve a high quality lysate every single time.
Keep sample as cool as possible
Protein degradation is a common yet unfortunate result of sample mishandling. Protein degradation results from disruption of the cell membrane, which releases proteases that can degrade the molecule of interest (oftentimes a protein). Samples need to be kept cool at all times to prevent protease activity. This may mean keeping the sample on ice while it is being used, precooling glassware and materials that the sample will come into contact with, and working quickly. Additionally, avoid freeze-thawing multiple times, as that will also degrade the sample over time. (1)
Select a method that complements the molecule of interest
Multiple cell lysis methods are available to researchers; determining your molecule of interest and the cell type it hails from will allow you to make an informed choice of lysis method. The two major classes of method are physical and detergent-based disruption. Physical disruption incorporates mechanical disruption, liquid homogenization, and grinding, while detergent-based disruption uses different chemical combinations to derive solutions that may be ionic, nonionic, or zwitterionic. It is important to research each method to find the one that will provide you with a high-yield, high-quality lysate.
Use protease inhibitors
As mentioned above, proteases can quickly degrade your protein of interest if the sample is not handled properly. By binding to proteases, protease inhibitors will prevent their destructive actions. Because multiple types of proteases are found in a single cell, use of a protease inhibitor cocktail will target many more culprit enzymes than a single protease inhibitor solution. Protease inhibitors can be purchased both individually or as cocktails.
Pion: The Homogenizer Advantage
Use of flexible and high quality lysis equipment will provide numerous benefits for both production laboratories and consumers alike. And there are plenty of companies on the market to select your equipment from. However, the lysate can be of higher quality and more even consistency when run through top-shelf equipment, most frequently in the form of a homogenizer. Pion's BEE brand technology is trusted by researchers and lab managers around the world. We deliver an array of key benefits, such as production of nano/micro emulsions and dispersions and lipids and suspensions; these can be used for applications such as injectables, targeted drug delivery, inhalants, time release, anesthetics, and importantly, vaccinations.
In addition, we have extensive experience in the challenges that our customers face as they transition from concept, through to R&D, clinical trials, all-important FDA approval and finally, to manufacturing.
Learn more by visiting our products page.