siRNA / miRNA gene silencing Mouse Embryonic stem cells

Get tips on using ON-TARGETplus Mouse Cdh5 (12562) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - MS1 Cdh5

Get tips on using ON-TARGETplus Mouse Cd84 (12523) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - M210B4 Cd84

Get tips on using ON-TARGETplus Mouse Tead4 (21679) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - C2C12 Tead4

Get tips on using ON-TARGETplus Mouse Tead2 (21677) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - C2C12 Tead2

Get tips on using ON-TARGETplus Mouse Tead1 (21676) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - C2C12 Tead1

Get tips on using ON-TARGETplus Mouse Sirt2 (64383) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - RAW264.7 Sirt2

Get tips on using ON-TARGETplus Mouse Bnip3 (12176) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - RAW264.7 BNIP3

Get tips on using ON-TARGETplus Mouse Rheb (19744) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - RAW264.7 Rheb

The RNA interference (RNAi) is used to inhibit gene expression or translation, by neutralizing targeted mRNA molecules. Two types of RNA molecules such as microRNA (miRNA) and small interfering RNA (siRNA) play a central role in RNAi. Few points have to considered to increase the transfection efficiency of siRNA. Always use healthy, actively dividing cells to maximize transfection efficiency. The confluency of cells should be between 50-70%. Always use the most appropriate siRNA concentration to avoid off-target effects and unwanted toxic side effects. Positive and negative controls should be used for each and every experiment to determine transfection efficiency.

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.