Hyaluronic acid (also called HA or Hyaluronan) is a glycosaminoglycan with elevated viscosity, enabling tissues (eye, skin, joint and synovial fluid…) to resist to physical and mechanical constraints (torsion, flexion…). Over time, when HA production declines, tissues progressively lose these tensile properties, leading to wrinkles and fold, weak re-epithelisation and age-related troubles. But HA is also involved in many other chronic and cancer-related diseases. In this post, we’ll review one of the most popular HA quantification assays (ELISA test), known to be highly sensitive and robust, appreciated by researchers involved in cosmetology and drug discovery.
Gel filtration (GF), also referred as Size Exclusion Chromatography (SEC), plays a key role in the high quality purification of enzymes, polysaccharides, nucleic acids, proteins and other biological macromolecules. Gel filtration is the simplest and mildest of all chromatography techniques to separate biomolecules on the basis of difference in size. Nevertheless, the list of available columns is quite awesome and the characteristics of each of them are very different. Also, each characteristic highly influences the quality of the final purified product with important consequences in downstream applications.
I’ve put together a simple guide to help you find your GF system, together with some tips and tricks based on my experience when producing recombinant proteins and antibodies for our clients involved in the early R&D stages through to the latest phases of the bio-production flow.
Download your copy of the guide “Gel Filtration – which column should you choose for your size exclusion purification?“
If you have any questions or if you need some help, leave a message below to get in touch with our lab experts!
The sensitivity and specificity of the primary and secondary antibodies used together with the IHC procedure used, are critical to avoid biased results. Several factors can cause false-positive or false-negative data, so they should all be verified as much as possible for each experimental set-up.
- Detection of the antigen of interest by the primary antibody
- Detection of the primary antibody by secondary antibodies
- Tissue preparation
Here, let’s look at 3 tips that will be of help to improve your IHC data.
Antibody-based techniques are widely used in Life Science laboratories. Antibody purification is often required to raise purity yield of antibody production batches or to reach publication-grade data. In this post, take advantage of some technical tips and troubleshooting points to improve your antibody purification.
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