siRNA / miRNA gene silencing Rat Schwann cells

Get tips on using Lipofectamine® 2000 Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - A549 & LTEP-a-2 Lipofectamine

Get tips on using TaqMan® MicroRNA Reverse Transcription Kit to perform siRNA / RNAi /miRNA transfection Mouse - Glomerular Mesangial cells polymer / lipid

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human pulmonary artery endothelial cells

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 Human/Mouse/Rat Total HSP70/HSPA1A DuoSet IC ELISA to perform ELISA Rat - HSP70

Get tips on using Rat Kidney injury molecule 1,Kim-1 ELISA Kit to perform ELISA Rat - KIM-1

Get tips on using Rat TIM-1/KIM-1/HAVCR Quantikine ELISA Kit to perform ELISA Rat - KIM-1

Get tips on using Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit to perform ELISA Rat - IGF-I

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Cells - primary human pulmonary artery smooth muscle cells

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.