A protein purification workflow typically involves many, often intricate, steps. While having the right reagents is critical to success, the right lab equipment and supplies can help make your workflow more simple, more efficient, and more effective. In this blog, we take you through some of the major steps commonly undertaken in a protein purification workflow, highlighting where the right lab equipment can help make your lab work easier!
1. Break Free with Cell Lysis
The first step is likely the lysing of cells to access their contents – including those all-important proteins – be they native or recombinant. There are several methods to choose from however, if you opt for a chemical approach, prepping your lysing solutions is often first on the agenda.
Accurate weighing of minute amounts of reagents, like lysozyme and DNase, can be tedious and requires a huge amount of concentration. Typical weighing approaches using plastic weighing boats and metal spatulas often result in much of the reagent being left behind as a residue, leading to inaccurate concentrations and variable results. Using a vibrating spatula can remove much of the hassle; giving you more control over the transfer from spatula to weighing dish and allowing for minute amounts of powder to be accurately weighed.
Next, it’s time to make sure your solutes successfully dissolve and your lysing solutions are properly mixed. You may reach for any magnetic stirrer you have to hand, but the size, shape and materials (coating and magnet) of the stirrer can all affect the stirring results. Our autoclavable Saturn SpinBar, for example, has a large sphere at the center that lifts the stirring bar ends while spinning, acting to reduce drag.
When it comes to lysing your cells, it’s important to keep your tubes organized and accessible. Having tubes of different sizes can make this difficult but the SP Bel-Art conical tube holder sets up in just seconds to hold either a 15 mL or a 50 mL conical tube.
2. Isolate Those Proteins!
Following cell lysis, the next step is to isolate your proteins. Common protein isolation and purification methods involve either chromatography, magnetic beads, or a range of other techniques. The good thing is, no matter which one you choose for your application, we’ve got you covered with the right supplies. Sometimes you’ll need to isolate several proteins at once; maybe you’ve generated wildtype and mutant proteins, or proteins with and without tags. Something as simple as finding the right rack for your application can not only help you stay organized but also help you effectively and efficiently carry out your protocol.
Magnetic bead purification is a popular purification method due to its relative simplicity and the ability to be performed at the bench. Whether you want to perform your assays in 96-well format or in 50 mL conical tubes, we offer a selection of powerful magnetic racks that make magnetic bead protein purification even simpler. In addition, clever design additions can help you tailor your protocols. For example, a spring plunger on tube sleeves of 1.5 – 50 mL models allows beads to be sequestered at various heights up the tube wall – preventing the inadvertent loss of critical material when removing supernatants.
Finally, when it comes to that all-important buffer exchange, dialysis is key! If you’re looking to remove imidazole or other interfering buffer components, we offer a range of tools – including dialysis equilibrium cells, tube funnels, and clip holders – to help streamline your workflow.
3. Don’t Just Trust. Verify.
Although still used as a protein purification method, more often than not, electrophoresis is used in protein studies as a verification technique. An electric field separates protein molecules in a solvent based on their charge and size, providing a simple, quick, and sensitive analytical tool. The most common electrophoretic technique used in protein purification workflows is the Western Blot – a method that is notoriously laborious and error-prone. By using the right lab supplies such as fixing trays and staining boxes that facilitate minimal gel handling, and orbital vibrating platforms that provide thorough yet gentle mixing of gels, you can increase workflow efficiency for more consistent results.
4. The Final Countdown – Streamlining Protein Assays
Once you’ve purified your proteins, you’ll be looking to assess them, for which there is an endless list of assays! Whether it’s protein detection or activity studies, SP Bel-Art stock an exhaustive list of lab equipment and supplies to match this diversity.
For example, one of the biggest challenges you may encounter during any protein assay is pipetting inconsistency and data variability. Using a repeating pipettor can help to improve reproducibility with precise, accurate, and reliable repetitive pipetting of volumes from 1 – 5000 µL.
If you’re investigating protein uptake, then radiolabeling can be a useful tool to track success, as it can relay vital information in real-time. However, radiolabeling proteins exposes the user to radioactive isotopes. Using the Beta Shield and Beta Box can help reduce your exposure while also providing excellent clarity.
From upstream cell lysis to downstream protein analysis, SP Industries stock a broad portfolio of equipment and supplies that will help simplify your protein purification workflow.