shRNA gene silencing Human Neuroblastoma cells (SH-SY5Y)

Get tips on using Gabpb1 siRNA to perform siRNA / miRNA gene silencing Rat - Schwann cells Nrf2

Get tips on using Ccr2 siRNA to perform siRNA / miRNA gene silencing Rat - Glial cells CCR2

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using N-WASP siRNA (h) to perform siRNA / miRNA gene silencing Human - T47-D N-WASP

Get tips on using VEGF-D siRNA (h) to perform siRNA / miRNA gene silencing Human - Caki-2 VEGF-D

Get tips on using EPAS-1 siRNA (h) to perform siRNA / miRNA gene silencing Human - HeLa EPAS-1 Lipid

Get tips on using IL-8 siRNA (h) to perform siRNA / miRNA gene silencing Human - HUVEC IL-8 Lipid

Get tips on using SignalSilence® SAPK/JNK siRNA to perform siRNA / miRNA gene silencing Human - KGN SAPK/JNK

Get tips on using siRNA ATX-1 or ENPP2 to perform siRNA / miRNA gene silencing Human - A2780 ATX-1

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 the 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.