Is hyaluronic acid content analysis by carbazole assay applicable to cell experiments?
I'm looking for hyaluronic content analysis not use ELISA kit.
Hello, Gyuseon!
Measurements of hyaluronan concentrations
The cell medium was collected to measure extracellular HA; then cells were treated with trypsin to collect pericellular HA; finally they were lysed with 0.1 M Tris-HCl buffer (pH 7.4) containing 10 mM CaCl2 to measure intracellular or intra-tissular HA. The medium and extracts were incubated overnight with 500 mU/ml Pronase (Sigma-Aldrich, Bornem, Belgium) at 55°C. HA concentration was quantified using Hyaluronan DuoSet kit (R&D systems, Abingdon, UK). Total cell protein concentration was measured using Pierce Protein Assay (ThermoScientific, Illinois, USA).
Article Hyaluronan Metabolism in Human Keratinocytes and Atopic Derm...
Article A simple method for hyaluronic acid quantification in culture broth
Hi Gyuseon Hwang!
I think it is important for you to identify whether you want to measure glycosaminoglycans (GAGs) in general or specifically hyaluronan/hyaluronic acid (HA).
Quantifying HA without ELISA is quite tricky and ELISA might be the simplest way of doing it. Other semi-quantitative ways include Particle Exclusion Assays, and staining with biotinylated HA-binding protein.
What Alexandr Chernov has talked about is a great way to isolate and measure HA. You might want to consider using Benzonases as well.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA