siRNA / RNAi /miRNA transfection Rat IEC-6

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.

Get tips on using HiPerFect Transfection Reagent to perform siRNA / RNAi /miRNA transfection Rat - IEC Cationic lipid based

I would like to regulate the expression of a gene and in order to do that, I have purchased specific siRNA. After optimizing my transfection protocol and using electroporation I have achieved a 60-70% reduction of the gene of interest. However, I cannot observe a significant reduction of mRNA expression but only a reduction of protein. What might be the problem? Could the problem be in my cell treatment method?

Get tips on using ON-TARGETplus Rat Smad2 (29357) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - IEC-6 Smad2

Get tips on using ON-TARGETplus Rat Smad3 (25631) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - IEC-6 Smad3

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.

Get tips on using HuR siRNA to perform siRNA / miRNA gene silencing Rat - IEC-6 HuR Lipid

Get tips on using - to perform siRNA / miRNA gene silencing Rat - IEC-6