Wound healing assay cell type rat

Get tips on using PE Rat Anti-Mouse CD184 to perform Flow cytometry Anti-bodies Mouse - CD184/CXCR4

Get tips on using PE Rat anti-Mouse CD146 to perform Flow cytometry Anti-bodies Mouse - CD146/MCAM

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.

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.

Transfection is a powerful technique that enables the study of the function of genes and gene products in cells. Based on the nature of experiments, we may need a stable DNA transfection in cells for persistent gain-of-function or loss-of-function of the target gene. For stable transfection, integration of a DNA vector into the chromosome is crucial which requires selective screening and clonal isolation. By carefully selecting a viral delivery system and related reagents we can ensure safe and highly-efficient delivery of expression constructs for high-level constitutive or inducible expression in any mammalian cell type.

Get tips on using Thiazolyl Blue Tetrazolium Bromide to perform Cell cytotoxicity / Proliferation assay cell type - HT22 mouse hippocampal cells

Get tips on using ON-TARGETplus Rat Prkaa1 (65248) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - UMR‐106 Ampkα1/Prkaa1

Get tips on using ON-TARGETplus Rat Prkaa2 (78975) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - UMR‐106 Ampkα2/Prkaa2

Get tips on using ON-TARGETplus Rat Arhgef6 (363509) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 α-Pix/Arhgef6

Get tips on using ON-TARGETplus Rat Arhgef7 (114559) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 β-Pix/Arhgef7