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Sucrose Gradient Centrifugation

Background Our Service Highlights FAQs Published Data Resource

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 Assembly defect result from the lack of free 5S rRNA. (Huang, Shijie, et al., 2020)Fig. 1 The lack of free 5S rRNA results in assembly defects.¹

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.

FAQs

Q1: What is Sucrose Gradient Centrifugation and its significance in research?

A: Sucrose Gradient Centrifugation is a technique that separates molecules based on their density using a sucrose density gradient. This method is crucial for monitoring active translation, a process integral to understanding cellular function and disease mechanisms. It allows for the isolation of ribosomes and mRNA complexes, thereby facilitating the study of translation misregulation, which can lead to various diseases. Its ability to characterize subcellular particles makes it valuable in biological research and analysis.

Q2: How does Creative Biolabs support researchers with Sucrose Gradient Centrifugation?

A: Creative Biolabs offers customized ribosome isolation and analysis services using Sucrose Gradient Centrifugation, tailored to meet diverse experimental goals. Our team of experts provides professional solutions and advanced technology to enhance research outcomes. We assist in designing optimal extraction protocols, overcoming the technique's limitations, and ensuring high-quality results for downstream analysis. Researchers are encouraged to contact us for customized service inquiries.

Q3: Can you provide guidance on how I can delve further into the range of services offered by Creative Biolabs in relation to ribosomes?

A: We encourage you to connect with our team of specialists for more detailed information. For those with specific requirements, there is an option to request a personalized quote. If you prefer a more general inquiry, simply reach out to us and we will design a solution tailored to your unique needs.

Published Data

Purification of eukaryotic ribosomes by isopycnic centrifugation in sucrose

ANALYTICAL BIOCHEMISTRY

Authors: P. Nieuwenhuysen, H. Slegers

Abstract:

A simple and inexpensive procedure for the isolation and purification of ribosomes from eukaryotes is described. The method avoids pelleting of ribosomes at high centrifugal forces and involves isopyenic centrifugation of the post-mitochrondrial supernatant in sucrose, precipitation of ribosomes with 10% polyethylene glycol, and zonal sucrose gradient centrifugation. The ribosomes obtained in this way are very pure and thus especially suited for the measurements of physical properties. The isopycnic centrifugation can also be used for the purification of other macromolecules and is only limited by a maximum density of sucrose of 1.40 g/cm3 obtained at the bottom of the centrifugation tubes.

Analysis by linear 5-20% sucrose gradient centrifugation of different density regions of a preparative sucrose isopycnic gradient. (Nieuwenhuysen, P., et al., 1978)Analysis by linear 5-20% sucrose gradient centrifugation of different density regions of a preparative sucrose isopycnic gradient. (Nieuwenhuysen, P., et al., 1978)

Resource

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. Huang, Shijie, et al. "Ribosome engineering reveals the importance of 5S rRNA autonomy for ribosome assembly." Nature Communications 11.1 (2020): 2900.
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