Understanding the principle behind the SDS-PAGE Analysis
Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis, commonly known as SDS-PAGE analysis, is a type of gel electrophoresis and an important technique for protein analysis. Gel electrophoresis is based on the principle of gel loading the samples and allowing the individual proteins to separate depending on how fast they migrate through the gel under the influence of an electrical field.
The Advantages of Gel Electrophoresis
• Gel electrophoresis allows the separation, identification, and visualization of proteins.
• Using SDS-PAGE analysis, you can estimate a sample’s purity and molecular weight.
• Via SDS-PAGE analysis you can quantify the amount of a protein in the sample.
• Gel electrophoresis is a primary step in various other techniques such as immunoblotting.
Easy gel loading and reliable outcomes of SDS-PAGE analysis with CAPP
Here at CAPP, we have optimized our products to meet the accuracy and sensitivity required for SDS-PAGE analysis protocol. With a combination of precision and ease, we ensure uniform measurements, comfortable gel loading and optimal results for your gel electrophoresis runs.
A successful SDS-PAGE analysis begins with successful sample preparation.
To prepare for gel loading and beginning your gel electrophoresis, you should begin by denaturing your protein sample. Mix adequate amounts of protein solution and SDS-PAGE sample buffer in 1.5mL CAPP Expell Secure Microcentrifuge Tubes. The advantages of using Expell Secure start with their leak-proof design that ensures your protection while handling hazardous chemicals used during SDS-PAGE analysis. Expell Secure tube are also designed with a polished interior to minimize sample binding to their inner surface, thus contributing to consistent gel electrophoresis results.
For high accuracy measurements and easier pipetting, we recommend you use CAPP ecopipette. The ecopipette, ecofriendly range, with variable volumes (2-20µl) and universal tip compatibility, was developed from biodegradable material, to save your bench space from crowding with all those fixed volume pipettes, without compromising the accuracy of your gel loading and SDS-PAGE analysis.
For effective sample denaturation for SDS-PAGE analysis, it is best to heat the mixture in a water bath placed over CRS-22H CAPPRondo Hotplate Stirrer for gradual heating, or the more compact, programable, CRS-21H CAPPRondo Magnetic Hotplate Stirrer, to ensure a smooth gel electrophoresis run upon gel loading.
Gel electrophoresis next step: Take your samples for a final spin down!
We are all aware of the danger of an imbalanced centrifuge, this is why we have focused our minds at CAPP on developing the imbalance detection feature to ensure safe centrifugation while you prepare for the gel loading phase of your gel electrophoresis.
You can either choose the fixed speed CAPPRondo Microcentrifuge CR-68, the CR-68X centrifuge with variable speed or the higher throughput CAPPRondo High-Speed Mini-Centrifuge CR-1512 equipped with a rotor accommodating up to 12 microcentrifuge tubes saving you valuable time while preparing for SDS-PAGE analysis or agarose gel electrophoresis.
Gel quality is a vital factor in gel electrophoresis
Pre-casted commercial gels are not always the best option for SDS-PAG analysis. In order to formulate your own gel, start by finding your optimal gel concentration (a rule of thumb for gel electrophoresis: the higher the concentration, the smaller the pores). Add the acrylamide, bis and water into 15 or 50 mL BluCAPP centrifuge tubes. BluCAPP’s leak-proof cap is designed with large ridges to provide you with an optimal, safe grip while preparing for gel electrophoresis. Moreover, it also supports one-handed operations by featuring a 3-4 turn closure, allowing a smoother and faster SDS-PAGE analysis.
A homogenous gel texture is crucial for SDS-PAGE analysis, to flawlessly homogenize the components, CAPPRondo Magnetic Stirrer CRS-15X would be an excellent choice! It has a compact structure with adjustable speed, pulse mode and a spill-resisting surface ideal for SDS-PAGE analysis and gel loading preparation.
Did you know casted polyacrylamide gel can be stored for several weeks?
Many researchers using SDS-PAGE analysis find it convenient to perp multiple gel casts for later gel loading. However, the prolonged continuous pipetting while casting and gel loading can subject scientists to wrist injuries!
At CAPP, we have solved this issue with CAPPSolo single channel pipette (5μL to 10mL), with a light design lowering the risk of wrist injury, it provides swift gel application and easy gel loading for a comfortable gel electrophoresis practice.
Finish gel loading and get ready to run!
Gel loading the samples into the wells for gel electrophoresis can seem like a daunting task. Samples such as protein or nucleic acids have high viscosity and low surface tension, making them prone to bubble formation, as well as leaving remnants inside the tips during gel loading. Such issues lead to sample dispersion and inconsistent results in SDS-PAGE analysis.
To prevent this, CAPP has come up with ExpellPlus, Premium, Low Retention Tips. Their low retention surface prevents the formation of bubbles within the sample, and minimizes sample loss during gel loading, giving your SDS-PAGE analysis clean bands with reproducible results.
Concerned about shooting the tip through the wells during gel loading?
For many researchers and students new to SDS-PAGE analysis and gel loading, preserving the wells from breakage can be challenging, but not anymore! ExpellPlus tips are produced with various lengths of the same handling volume, allowing comfortable gel loading while eliminating your fear of breaking the gel while performing gel electrophoresis.
Visualizing the results of SDS-PAGE analysis
The gel loading and gel electrophoresis are complete! To visualize the bands, soak the gel in a mixture containing your designated dye, follow this by washing to remove any excess dye. The finishing of SDS-PAGE analysis requires gentle shaking and even gel coating.
The ultimate choice here is the CAPPRondo line. The CR3-08X 3D Shaker features 3 dimensional movements optimal for evenly coating your gel, and if you prefer orbital motion, you can try CAPPRondo Platform Shaker CRP-3X with multiple programs and interchangeable attachments, such as the slip resisting tray CRP-D for an extra sense of stability while performing gel electrophoresis.
Inspired by the researchers’ needs & their everyday lab life
With over 30 years of developing top quality laboratory equipment, CAPP is now here to take you on a swift gel loading experience and a successful gel electrophoresis practice. At affordable prices and without any compromises on the quality. CAPP instruments require minimal maintenance and most of our liquid handling utensils are autoclavable and feature universal compatibility, making our products the most comfortable and cost-effective option for your SDS-PAGE analysis protocol.
Get your hands on CAPP’s top quality equipment for gel loading and SDS-PAGE analysis NOW!
All CAPP products used for this gel loading & SDS-PAGE analysis protocol are available for a FREE trial, wherever you are, via our worldwide distributor network. All you have to do is click here and we will arrange for test equipment to arrive at your lab in time for your gel electrophoresis experiment, FREE of charge!