crispr-mouse-activation-3t3-l1-c-ebp

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Get tips on using eBioscience™ Annexin V-FITC Apoptosis Detection Kit to perform Apoptosis assay cell type - MCF-7

Products Thermo Fisher Scientific eBioscience™ Annexin V-FITC Apoptosis Detection Kit

Get tips on using B7-H4 Monoclonal Antibody (188), PE, eBioscience™ to perform Flow cytometry Anti-bodies Human - B7-H4

Products eBioscience B7-H4 Monoclonal Antibody (188), PE, eBioscience™

Get tips on using eBioscience™ Annexin V-FITC Apoptosis Detection Kit to perform Apoptosis assay cell type - SMMC-7721, HEPG2

Products Thermo Fisher Scientific eBioscience™ Annexin V-FITC Apoptosis Detection Kit

Get tips on using FOXP3 Monoclonal Antibody (PCH101), Alexa Fluor 700, eBioscience™ to perform Flow cytometry Anti-bodies Human - FOXP3

Products eBioscience FOXP3 Monoclonal Antibody (PCH101), Alexa Fluor 700, eBioscience™

Get tips on using FOXP3 Monoclonal Antibody (eBio7979), eBioscience™ to perform Western blotting FOXP3

Products Thermo Fisher Scientific FOXP3 Monoclonal Antibody (eBio7979), eBioscience™

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.

Proteins Protein Expression Prokaryotic cells E. coli 2-epimerase

Get tips on using Hoechst 33258 to perform DNA quantification Human - MDA-MB-231

Products Thermo Fisher Scientific Hoechst 33258

The challenge in isolating RNA from S. aureus cells is the disruption of the cell wall. A lot of protocols employ enzymatic digestion (pretreatment) which may affect gene expression patterns of certain genes. Therefore physical disruption using beads is considered to be the better alternative.

RNA RNA isolation / purification Bacteria Gram positive Staphylococcus aureus

The biggest problem in isolating RNA from gram-positive bacteria is the disruption of the cell wall. A lot of protocols employ enzymatic digestion (pretreatment) which may affect gene expression patterns of certain genes. Therefore physical disruption using beads can be a best alternative.

RNA RNA isolation / purification Bacteria Gram positive Clostridium difficile

The biggest problem in isolating RNA from gram-positive bacteria is the disruption of the cell wall. A lot of protocols employ enzymatic digestion (pretreatment) which may affect gene expression patterns of certain genes. Therefore physical disruption using beads can be a best alternative.

RNA RNA isolation / purification Bacteria Gram positive Clostridium tetani

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