Get tips on using Lipofectamine® 2000 Transfection Reagent to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells IGFBP1 (Insuline-like growth factor binding protein-1) Lipid
Get tips on using Galacto-Star™ β-Galactosidase Reporter Gene Assay System for Mammalian Cells to perform Reporter gene assay β-galactosidase substrates - MCF-7 human breast cancer
Get tips on using MISSION® pLKO.1-puro Non-Mammalian shRNA Control Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans Negative control (scrambled) lentiviral particles
Get tips on using STEMdiff™ Trilineage Differentiation Kit to perform Stem cell Differentiation media Differentiation of Human primed induced pluripotent stem cells (UMN PCBC16iPS) into naive pluripotent stem cells
Get tips on using OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) to perform ROS assay cell type - PLHC-1, SK-HEP-1, Hep3b, HepG2 human hepatocellular carcinoma
Get tips on using APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU to perform TUNEL assay cell type - A549, NCI-H460, H1299 human alveolar carcinoma
Get tips on using Pre-designed and validated siRNA against gene IGFBP1 to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells IGFBP1 (Insuline-like growth factor binding protein-1)
Get tips on using pET-28a-chBCO2 to perform Protein Expression Prokaryotic cells - E. coli chicken BCO2
I have tried to fabricate Liver organoids and would like to study the impact of FBS on healthy and tumor organoids. Since the compositions of FBS is unknown, do you recommend any alternatives like Human platelet lysate, etc?
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.
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