Site Directed Mutagenesis (SDM) Human Deletion MCF-7

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Get tips on using CellROX™ Deep Red Reagent, for oxidative stress detection to perform ROS assay cell type - human umbelical vein endothelial cells (HUVEC)

Products Thermo Fisher Scientific CellROX™ Deep Red Reagent, for oxidative stress detection

Get tips on using High Pure RNA Isolation Kit to perform RNA isolation / purification Cells - primary human aortic smooth muscle cells

Products Roche Lifesciences High Pure RNA Isolation Kit

Get tips on using Gibco™ DMEM, high glucose, GlutaMAX™ Supplement to perform Stem cell culture media Human Dental pulp stem cells (hDPSC)

Products Thermo Fisher Scientific Gibco™ DMEM, high glucose, GlutaMAX™ Supplement

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.

Cellular assays Autophagy assay cell type MEFs (mouse embryonic fibroblasts)

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

Proteins Western blotting Smad2

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

Proteins Western blotting Smad4

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

Proteins Western blotting Smad1

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

Proteins Western blotting Smooth muscle actin

Get tips on using Biotinylated Protein Ladder Detection Pack #7727 to perform Protein Ladder Immunofluorescence

Products Cell Signaling Technology Biotinylated Protein Ladder Detection Pack #7727

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human peripheral blood mononuclear cells

Products Thermo Fisher Scientific TRIzol Reagent

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