Sucrose Gradient Centrifugation

Sucrose density fractionation has long been considered the gold standard for monitoring active translation. Creative Biolabs is a leading global provider of ribosome isolation and analysis services. Our expert team provides the most professional solutions and advanced technology to promote your research.

Background

What is Sucrose Gradient Centrifugation?

Misregulation of translation can lead to a variety of diseases. The cellular mRNA translation process is inseparable from the ribosome. Actively translated mRNA is usually bound by several ribosomes and can be separated from small (40S) and large (60S) ribosomal subunits and 80S monomers by sucrose gradient centrifugation. Total RNA was isolated from different parts of the multimer gradient and detected by different methods.

Sucrose gradient centrifugation is considered a common and efficient method for characterizing subcellular particles. The technique separates targets based on a density gradient of sucrose. The main principle is that the sample rotates under centrifugal force, and the centrifugal force generated by the rotation of the sample causes the molecules to move in the medium. Different molecules move through the gradient until their density is the same as that of the surrounding medium. This allows molecules of different densities to separate at different layers, which can then be used in various recycling processes.

Fig.1 Schematic of a sucrose gradient fractionation of ribosomes. (Gerashchenko, 2017)

Applications of Sucrose Gradient Centrifugation

• This centrifugal method can be used to separate macromolecules such as DNA and RNA.
• This can also be used to analyze protein complexes, as well as to determine the size and density of other macromolecules.
• In addition, this method plays a significant role in the isolation of ribosomes as well as membrane components, centrosomes, and subcellular organelles.

Limitations of Sucrose Gradient Centrifugation

Although sucrose density fractionation plays an important role in the separation of ribosomes as well as other molecules, there are some disadvantages:

• Need special equipment, for example, it requires conditions such as ultracentrifuge, and gradient fractionation system.
• The procedure is time-consuming and does not allow multiple samples to be processed simultaneously.
• The samples will be diluted when added to the sucrose solution, therefore, more elaborate subsequent steps are required to obtain samples of sufficient quality for downstream experiments.
• The obtained samples are easily contaminated by macromolecules such as lipid rafts and pseudopolysomes.

Our Service

Creative Biolabs is a leader in ribosome isolation and analysis, utilizing sucrose gradient centrifugation—a method integral to our wide range of services aimed at advancing research through professional, state-of-the-art solutions. Dedicated to providing services that cater to varied research goals, we excel in fine-tuning extraction protocols to deliver unparalleled quality results, thereby facilitating your research journey. We encourage researchers from all corners of the globe to discover our bespoke services, meticulously crafted to fulfill your unique experimental requirements.

Highlights

• Expert Ribosome Isolation and Analysis

We offer leading-edge ribosome isolation and analysis services, leveraging our expert team's proficiency to deliver professional solutions that advance your research.

• Advanced Technology and Custom Solutions

Our application of advanced technology and commitment to customized service ensures that every project meets the unique experimental goals of our clients, from protocol design to final analysis.

• Versatile Applications

Our sucrose gradient centrifugation service is versatile, capable of separating and analyzing a wide range of macromolecules including DNA, RNA, protein complexes, and characterizing the size and density of these macromolecules. It is also instrumental in the isolation of ribosomes, membrane components, centrosomes, and various subcellular organelles.

• Comprehensive Support and Optimization

Acknowledging the complexities and potential limitations of sucrose gradient centrifugation, such as the need for special equipment and the time-intensive nature of the process, Creative Biolabs provides extensive support to navigate these challenges. We assist in designing optimal extraction protocols and offer solutions to achieve high-quality samples suitable for downstream experiments.

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Creative Biolabs provides customized services to global customers according to different experimental goals. We will help you design the optimal extraction protocols to achieve the best results. Please feel free to contact us.

Reference

1. Gerashchenko, M. V.; Gladyshev, V. N. Ribonuclease selection for ribosome profiling. Nucleic acids research. 2017, 45(2): e6-e6.