Get tips on using Stealth siRNA(m)_Atg5 to perform siRNA / miRNA gene silencing Mouse - MLO‐Y4 Atg5
Get tips on using Gibco™ StemPro™ hESC SFM to perform Stem cell Differentiation media hiPSCs or hESCs differentiation to Embryoid body (EB)
Get tips on using Stealth siRNA(r)_Mmp15 to perform siRNA / miRNA gene silencing Rat - C6 (rat glioma) mmp15
Get tips on using Stealth siRNA(r)_Ctnnb1 to perform siRNA / miRNA gene silencing Rat - NPC β-Catenin/Ctnnb1
Get tips on using Stealth siRNA(m) Atg16l2 to perform siRNA / miRNA gene silencing Mouse - Pancreatic Acinar cells Atg16l2
RNAi or RNA interference is a common method to suppress gene expression in vitro/in vivo by utilizing the inherent microRNA machinery, without introducing a total gene knockout. 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.
Get tips on using ON-TARGETplus Mouse Samhd1 siRNA to perform siRNA / miRNA gene silencing Mouse - BMDMs SAMHD1
Get tips on using ViralSEQ™ Quantitative Sf-Rhabdovirus Kit with PrepSEQ™ Nucleic Acid Sample Prep Kit to perform Cell Culture Contamination Detection Kit Virus
As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.
As autophagy is a multi-step process which includes not just the formation of autophagosomes, but most importantly, flux through the entire system, including the degradation upon fusion with lysosomes, which makes it quite challenging for detection. There are several methods for detection in mammalian cells, including immunoblotting analysis of LC3 and p62 and detection of autophagosome formation/maturation by fluorescence microscopy, Currently, there is no single “gold standard” for determining the autophagic activity that is applicable in every experimental context, hence it is recommended to go for the combined use of multiple methods to accurately assess the autophagic activity in any given biological setting.
Fill out your contact details and receive price quotes in your Inbox
Outsource experiment