RNA isolation / purification Cells immortalized

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 Rn_Rab7_5 FlexiTube siRNA(r) to perform siRNA / miRNA gene silencing Rat - NRVM( Rab7

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

Get tips on using Rn_LOC311846_1 FlexiTube siRNA to perform siRNA / miRNA gene silencing Rat - Astrocytes LRRC8A

Get tips on using Rn_Sod2_4 FlexiTube siRNA to perform siRNA / miRNA gene silencing Rat - PC12 SOD2

Get tips on using Rn_Pdcd8_1 FlexiTube siRNA to perform siRNA / miRNA gene silencing Rat - H9c2 AIF/Pdcd8

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

Get tips on using Rn_LOC312647_1_ ATG7 FlexiTube siRNA(r) to perform siRNA / miRNA gene silencing Rat - NRVM( ATG7

Get tips on using Rn_Ywhaz_4 FlexiTube siRNA to perform siRNA / miRNA gene silencing Rat - H9c2 14-3-3 f/Ywhaz

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.