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mRNA isolation

Introduction and inspiration of mRNA

There is a lot to learn from the mRNA (messenger RNA) that is present in cells, which makes up around 5% of the total RNA in cells. The presence of certain mRNA sequences can inform us about what proteins are most likely being translated at the moment so that biological processes can take place. mRNA has also emerged as a top vaccine for the novel coronavirus of 2019, also known as SARS-CoV-2. How can a messenger RNA be a vaccine you ask? mRNA works as a vaccine by providing our cells with the sequence to make the major protein found on SARS-CoV-2 called “spike” protein. Once our cells have made spike protein, our body will launch an immune response against it, and immune cells will make antibodies against the spike protein. To use and study mRNA, you must first perform mRNA isolation, which we will introduce in this article. 

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Protein purification system

Proteins are one of the four main macromolecules in biology. They have many crucial functions in all living organisms. Research laboratories all over the world study the many types of proteins to better understand the mechanisms of living things

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RNA purification protocol

RNA (ribonucleic acid) is found in all living things. It has several functions in cells including playing a role in transcription, translation, regulation and gene expression.

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Chemiluminescence examples

In this post we will cover in detail the definition of chemiluminescence definition and some chemiluminescence examples to get you familiar with the phenomenon and its uses. Chemiluminescence is defined by the release of light from a chemical reaction. This process occurs naturally in fireflies and several types of sea creatures such as jellyfish, in these cases it is also called bioluminescence. Glowsticks, the vibrant party favors, are also a form of chemiluminescence. 

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A general filtration process

Filtration is a simple technique used to separate solid particles from suspension in a liquid solution. There are many filtration methods available, but all are based on the same general principle: a heterogenous mixture is poured over a filter membrane. The filter membrane has pores of a particular size. Particles larger than the pores will be unable to pass through the membrane, while particles smaller than the pores will pass through unhindered. Additionally, all liquids will pass through. The final result of a filtration process is a collection of residue on the filtration membrane. This residue is therefore effectively separated from the rest of the mixture that passed through the membrane.

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Cell sorting techniques

Fundamental research often involves the study of isolated cell populations. It is these enriched populations that enable researchers to make new discoveries about cell function, signaling, gene expression, fate decisions, and much more. Techniques for the rapid and accurate enrichment of target cell populations are an area of great interest. Cell sorting techniques fall into two general categories: bulk sorting and single cell sorting. In bulk cell sorting all of the target cells are collected in one sweep, whereas in single cell sorting every cell is individually analyzed. There are multiple methods of bulk cell sorting: filtration, centrifugation, and magnetic cell sorting. The main single cell sorting method is flow cytometry or fluorescence activated cell sorting. While cell sorting can be very accurate, it is hard to say that a sorted cell population is “pure”. Instead, the collected population is referred to as “enriched”. In general, single cell sorting results in highly enriched cell populations that are more homogeneous than those obtained via bulk sorting methods.

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Direct Elisa and Indirect Elisa

A couple of months ago we described the sandwich elisa. Here we will discuss the other two main types of elisas—indirect and direct. Elisa is an acronym for enzyme-linked immunosorbant assay. The purpose of any elisa is to detect the presence of an antigen or antibody of interest. The indirect and direct elisa differ from the sandwich elisa because the antigen of interest is bound directly to the plate rather than a capture antibody. In either case, the key component is an enzyme-linked detection antibody. The enzyme is either colorimetric or chemiluminiscent. Chemiluminiscent enzymes are popular because they are easily read by a luminometer plate reader, making the process easy and highly quantitative.

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Enzymes in Industry

Enzymes play a surprisingly important role in modern industry, and are essential to the production of more commercial products than one would initially consider. Enzymes are proteins that speed up reactions and improve yield by increasing available precursors for downstream reactions. Perhaps the most obvious use for enzymes in industryisthe production of cheese, bread, and alcohol. In these traditional applications the enzymes are part of microbial machinery such as bacteria or yeast. Over time scientists have been able to isolate specific enzymes and to understand their catalytic functions well enough to incorporate them with or without their microbial hosts into a wide variety of somewhat surprising situations. For example, enzymes are used in the production of textiles, detergents, biofuels, and pharmaceutical products. Large quantities of desired enzymes are required for these applications, and they need to be available in the purest form possible. The purity of enzymes in industry is particular important for pharmaceutical applications where the products as well as the process are susceptible to review and control by regulatory associations. Batches of enzymes in industry undergo regular process validation to ensure batch-to-batch consistency.

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The industrial centrifuge

The industrial centrifuge plays an integral role in the production of more things than one would initially expect. It is a commonly used tool in the food and agricultural sector, At pharmaceutical and biotechnology companies, for environmental management, and in the chemical industry. The word industry conjures up images of combination and creation—adding materials together to produce a final product. However, the separation of materials is just as important as the combination of materials. We can't create a new product until we have pure reactants to work with. This is especially important in the pharmaceutical and biotechnological realms, where reactant purity is essential to the production of a product that is safe for human consumption. This is where the centrifuge comes in. The centrifuge is used to separate heterogeneous mixtures into components varying by density.

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Elution Buffer

An elution buffer plays an essential role in every immunoprecipitation protocol or assay that requires the release of a target antigen from a capture antibody. Elution buffers are necessary in protocols utilizing a stationary affinity column, and are also required in protocols using mobile solid supports in solution.

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