Site Directed Mutagenesis (SDM) Rat Deletion H9C2

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.

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 In Situ Cell Proliferation Kit, FLUOS to perform Cell cytotoxicity / Proliferation assay cell type - HUVEC

Get tips on using siGENOME PCNT siRNA to perform siRNA / miRNA gene silencing Human - HEK293 PCNT

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Get tips on using siGENOME Human MINK1 siRNA to perform siRNA / miRNA gene silencing Human - RMS MINK

Get tips on using siGENOME Human MAP4K2 siRNA to perform siRNA / miRNA gene silencing Human - RMS MAP4K2

Get tips on using siGENOME Human PAK1 siRNA to perform siRNA / miRNA gene silencing Human - HeLa PAK1

Get tips on using siGENOME Mouse Alox12 siRNA to perform siRNA / miRNA gene silencing Mouse - B16-F10 12-Lox/ALOX12

Get tips on using siGENOME Human FTO (79068) siRNA - Individual to perform siRNA / miRNA gene silencing Human - SHSY5Y FTO