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.
Get tips on using Anti-LC3B antibody produced in rabbit to perform Autophagy assay cell type - Mouse lung tissue
Get tips on using Anti-LC3B antibody produced in rabbit to perform Autophagy assay cell type - Mouse embryonic fibroblasts
Get tips on using Anti-phospho-Histone H2A.X (Ser139) Antibody to perform TissueFAxs phospho-Histone H2A.X (Ser139) - Mouse Human
Get tips on using Anti-phospho-Histone H2A.X (Ser139) Antibody to perform Immunofluorscence phospho-Histone H2A.X (Ser139) - Mouse Human
Get tips on using Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA Kit to perform ELISA Rat - TGF-beta 1
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 SurePrint G3 Mouse Exon 4x180K Microarray Kit (165,984 Exon probes) to perform Microarray Mice - Cochlea Expression array (labelled)
Get tips on using anti-Mucin 6, Clone: CLH5, Invitrogen™ to perform Immunohistochemistry Human - MUC-6
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