DNA isolation / purification Cells Immortalized cell lines

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Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase

Cellular assays Reporter gene assay luciferase vascular smooth muscle cells (VSMC)

Get tips on using T-PER™ Tissue Protein Extraction Reagent to perform Protein isolation Mammalian cells - Rat_Liver

Products Thermo Fisher Scientific T-PER™ Tissue Protein Extraction Reagent

Get tips on using T-PER™ Tissue Protein Extraction Reagent to perform Protein isolation Mammalian cells - Huh7

Products Thermo Fisher Scientific T-PER™ Tissue Protein Extraction Reagent

Get tips on using M-PER™ Mammalian Protein Extraction Reagent to perform Protein isolation Mammalian cells - HUVEC

Products Thermo Fisher Scientific M-PER™ Mammalian Protein Extraction Reagent

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 DAOY

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 Daudi

Get tips on using MagniSort™ Human pan-Monocyte Enrichment Kit to perform Cell Isolation Monocyte

Products Thermo Fisher Scientific MagniSort™ Human pan-Monocyte Enrichment Kit

Get tips on using Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 HER2 leader peptide

Products Thermo Fisher Scientific Expi293™ Expression System Kit

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - adipose stem cells

Products Cell Signaling Technology Senescence β-Galactosidase Staining Kit - Cell Signaling

Contamination can affect cell characteristics, i.e., growth, metabolism, and morphology leading to unreliable and erroneous experimental data. Depending on the source of contaminants, one can detect contamination by using a light microscope, gram stain, isothermal amplification, or PCR. Bacteria and fungi can usually be identified by optical microscopy. Mycoplasma in cell cultures cannot be detected visually. Hence, these microbes can go unnoticed for long periods and are determined using dedicated assays. Early and rapid identification of contaminants is vital to detect, handle and prevent contamination for good cell-culture practices. However, detection and identification can be challenging and tricky based on usual visual identifications. Hence it is essential to use a standard contamination detection kit to detect and maintain best practices.

Cellular assays Cell Culture Contamination Detection Kit Mycoplasma

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