Protein Expression Eukaryotic cells A. thaliana

Get tips on using Mem-PER™ Plus Membrane Protein Extraction Kit to perform Protein isolation Mammalian cells - Mouse_Brown fat

Get tips on using PRO-PREP™ Protein Extraction Solution (C/T) to perform Protein isolation Mammalian cells - 3T3-L1

Get tips on using TAGZyme pQE Vector Set to perform Protein tag Expression of His-tagged proteins

miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time consuming, but provide a more permanent expression of RNAi in the cells, and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.

Get tips on using Qproteome Nuclear Protein Kit to perform Protein enrichment Soluble nuclear proteins

Get tips on using PRO-PREP™ Protein Extraction Solution (C/T) to perform Protein isolation Mammalian cells - Mouse Epididymal fat

Get tips on using SurePrint G3 Human Gene Expression 8x60K v2 Microarray Kit to perform Microarray Human - PCOS

Get tips on using Unstained Protein Standards to perform Protein Ladder Unstained

Get tips on using Prestained Protein Standards to perform Protein Ladder Prestained

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that no responses other than those related to the signaling pathway of interest. This can be achieved by selecting a highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzyme such as luciferase.