rna-isolation-purification-cells-immortalized-mda-mb-231

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 Rat / Mouse FGF-21 ELISA Kit to perform ELISA Rat - FGF-21

ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.

Get tips on using Fibroblast Growth Factor 21 Mouse/Rat ELISA to perform ELISA Rat - FGF-21

Get tips on using Mouse/Rat FGF-21 Quantikine ELISA Kit to perform ELISA Rat - FGF-21

Get tips on using Minimum Essential Media (MEM) to perform Mammalian cell culture media BHK-21

Get tips on using DMEM–Dulbecco's Modified Eagle Medium to perform Mammalian cell culture media BHK-21

Get tips on using HyClone RPMI 1640 media: Liquid to perform Mammalian cell culture media BHK-21

Get tips on using INTERFERin® to perform siRNA / RNAi /miRNA transfection Mouse - B16 Polymer / lipid