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Applications
- Protein Purification
- PCR Prep
- Chromatography
Protein Purification
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. Below are 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.
Cell Lysis
One of the first steps is the lysing of cells to access their proteins. Depending on the cell type, this may be done mechanically or chemically. Either way, the extracted proteins are delicate and care needs to be taken to make certain downstream steps are successful.
Chromatography
Protein chromatography separates proteins based on their properties. This allows for selection and isolation of the target proteins.
Dialysis
Dialysis is a separation technique that removes smaller, unwanted compounds from the solution of larger protein molecules. Molecules diffuse into either side of a selective membrane based on the molecular size.
Assay
Once interferences are removed by the purification steps, you are then able to quantify and detect the target protein with various techniques, such as such as fluorescence, spectrophotometry, or colorimetry.
Electrophoresis
Gel electrophoresis can be used to separate proteins for both analysis and purification. Once proteins have been separated, they can be used for downstream applications like enzymatic assays, further purification, immunoblotting, and elution and digestion for mass spectrometric analysis.
PCR Prep
Preparing for a successful PCR experiment requires optimal samples, reagents, and reaction protocols, but having the correct tools to prepare your PCR experiment can also improve the outcome of the reaction and increase the reliability of your results. These tools can provide you with better sanitation of the working environment, efficient thawing and mixing of samples and reagents, precise, accurate, and reliable repetitive pipetting of reagents and samples, and improved storage conditions.
From storage of your samples and reagents to thawing, mixing, and processing them, the SP Bel-Art™ portfolio of products can help you achieve successful and reliable PCR results every time. For example, SP Bel-Art™ LabMat™ bench liners can absorb and contain spills quickly, and wide-mouthed and trigger spray wash bottles can be used with bleach solution to decontaminate benches. For precise, accurate, and reliable dispensing of samples into a PCR prep, you can use a repeating pipettor, such as the SP Bel-Art™ Roxy M™ Repeating Pipettor and pipette into SP Bel-Art™ Magic Touch 2™ ice buckets and tube racks that can keep samples and reagents at stable temperatures throughout the process.
Sanitation
Lab benches, equipment, and devices can introduce contamination from previous DNA preparations. Because PCR is a sensitive and specific technique, attention must be given to sanitization, preparation, and organization prior to beginning PCR work.
Thawing
It is important to gently thaw samples and reagents that contain sensitive enzymes to ensure successful PCR.
Mixing
Proper mixing ensures that the chemicals needed for the PCR experiment support and contribute to the reaction.
Reagent and Sample Storage
Long-term storage of samples and enzyme reagents is done at sub-freezing temperatures.
Chromatography
Separating molecules by chromatography occurs across multiple branches of science and relies on successful fractionation. At each step of the process, having the optimal tools improves the chances of a well-defined separation of your sample components.
Preparation
When preparing the column, careful thought must be given to provide best sample separation. Consider sample polarity, size, and affinity. Preparation is dictated by what type of chromatography is used — analytical or preparative.
Running the Column
Columns must be set up properly for samples to run effectively. Factors like loading the column or making sure it is vertically straight could make a vast difference in sample collection.
Fraction Collection
Fraction collection varies, depending on how well you can visualize your sample on the column. Best practice dictates to collect many small fractions that can be pooled together once their purity is confirmed.
Clean Up
Properly cleaned glassware and plasticware allows for confidence in future analyses using the same methodology.
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Videos
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Bel-Art SP Scienceware Switch-Grid Test Tube Racks
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Bel-Art SP Scienceware Right-to-Know Safety-Vented Wash Bottles with GHS Labeling
Learn more about these wash bottles that are labeled to alert students to potential hazards and the need for additional protective equipment or clothing.
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About SP Bel-Art
SP Bel-Art brand labware includes a broad selection of innovative, tried-and-true product solutions that support research and production across diverse markets, including biopharma, life sciences, safety, academia, food and beverage, petrochemical, and clinical health.
The portfolio features over 3,500 labware products — from safety wash bottles to fume hoods — including H-B Instrument™ thermometers and timers, SP Bel-Art™ Poxygrid™ racks, Secador desiccators, Spinbar stirring bars, Sterileware scoops and spoons, and more. As part of the SP Industries family of brands, U.S.-based thermometer and hydrometer calibration services are also available.