ROS assay cell type

Get tips on using Bioanalyzer High Sensitivity DNA Analysis to perform DNA Damage Assay HT1080

Get tips on using Apoptotic DNA Ladder Isolation Kit to perform DNA Damage Assay U266

Get tips on using Renilla luciferase vector, pGL4.74 to perform Reporter gene assay luciferase - primary human endometrial stromal cells

Get tips on using Zombie Violet™ Fixable Viability Kit to perform Live / Dead assay mammalian cells - mouse microglia

Get tips on using Zombie UV™ Fixable Viability Kit to perform Live / Dead assay mammalian cells - mouse splenocytes

Get tips on using Zombie Fixable Viability™ Sampler Kit to perform Live / Dead assay mammalian cells - HEK 293

Get tips on using Live/Dead Double Staining Kit (Merck) to perform Live / Dead assay mammalian cells - THP-1

Get tips on using Senescence Cells Histochemical Staining Kit to perform Reporter gene assay β-galactosidase substrates - adipose stem cells

Get tips on using Senescence Cells Histochemical Staining Kit to perform Reporter gene assay β-galactosidase substrates - mouse embryonic fibroblasts

Protein expression refers to the techniques in which a protein of interest is synthesized, modified or regulated in cells. The blueprints for proteins are stored in DNA which is then transcribed to produce messenger RNA (mRNA). mRNA is then translated into protein. In prokaryotes, this process of mRNA translation occurs simultaneously with mRNA transcription. In eukaryotes, these two processes occur at separate times and in separate cellular regions (transcription in nucleus and translation in cytoplasm). Recombinant protein expression utilizes cellular machinery to generate proteins, instead of chemical synthesis of proteins as it is very complex. Proteins produced from such DNA templates are called recombinant proteins and DNA templates are simple to construct. Recombinant protein expression involves transfecting cells with a DNA vector that contains the template. The cultured cells can then transcribe and translate the desired protein. The cells can be lysed to extract the expressed protein for subsequent purification. Both prokaryotic and eukaryotic protein expression systems are widely used. The selection of the system depends on the type of protein, the requirements for functional activity and the desired yield. These expression systems include mammalian, insect, yeast, bacterial, algal and cell-free. Each of these has pros and cons. Mammalian expression systems can be used for transient or stable expression, with ultra high-yield protein expression. However, high yields are only possible in suspension cultures and more demanding culture conditions. Insect cultures are the same as mammalian, except that they can be used as both static and suspension cultures. These cultures also have demanding culture conditions and may also be time consuming. Yeast cultures can produce eukaryotic proteins and are scalable, with minimum culture requirements. Yeast cultures may require growth culture optimization. Bacterial cultures are simple, scalable and low cost, but these may require protein specific optimization and are not suitable for all mammalian proteins. Algal cultures are optimized for robust selection and expression, but these are less developed than other host platforms. Cell-free systems are open, free of any unnatural compounds, fast and simple. This system is however, not optimal for scaling up.