As a scientist/researcher, you can probably think of some lab techniques that have become second nature due to frequent use. However, what if you could make a few tweaks to an already existing technique to make it more efficient? This can translate to savings in both time and money, and importantly, increased yield and quality of lysate. Keep reading to learn about a few tips that will help make your cell rupture technique more efficient.
Decrease Number of Cycles
Although this strategy may only work with some homogenizers, those which may be impacted can see significant results. Most homogenizers require at least 3-4 passes to achieve a consistent and uniform product. However, some higher quality homogenizers can obtain the same results in just 1-2 passes. By decreasing the number of cycles, you can expect to save both reagents/resources and time.
Keep Sample as Cool as Possible
Protein degradation is a common yet unfortunate result of sample mishandling. When the cell membrane is disrupted, proteases that can degrade the molecule of interest, oftentimes a protein, are released. By keeping the sample cool, protease activity can be slowed and even prevented. You can take action by keeping the sample on ice during use, pre-cooling glassware, or working quickly; additionally, it is important to avoid repeated freeze-thawing, which can propagate the degradation process. (1)
In a study conducted by Ahmad-Raus et. al., researchers were interested in determining which high pressure homogenization conditions were most important to obtaining a high lysate yield. They found that, specifically among E. coli cells, high pressure (as opposed to medium or low pressure) had a huge impact on the lysate yield. It is therefore advisable to boost the pressure of your own technique for better time and product outcomes. (2)
Use Protease Inhibitors
Because proteases degrade proteins, your protein of interest may be at risk for denaturation if the sample is not handled properly. While cooling can help to slow the actions of proteases, as indicated above, protease inhibitors can completely rid the sample of these irksome molecules. By binding to proteases, protease inhibitors can prevent their destructive actions. As multiple types of proteases exist within a single cell, protein inhibitor cocktails that target many more culprit enzymes are recommended. Conveniently, most science supply companies provide protease inhibitors either individually or as pre-mixed cocktails.
Pion: The Homogenizer Advantage
One way to implement the above tips is to consider the equipment you are using for your cell disruption technique. As the preferred method, high pressure homogenization will save you precious time while still achieving a high yield of intracellular contents. Pion's BEE brand technology manufactures high pressure homogenizers that are trusted by researchers and lab managers around the world. We deliver an array of key benefits - importantly, cell disruption - but also production of nano/micro emulsions and dispersions and lipids and suspensions. These can be used for products across the pharmaceutical, biotechnology, food, and chemical industries.
Learn more about Pion's BEE brand homogenizers for cell disruption by contacting us today.