Protein Expression Eukaryotic cells Hi5

Get tips on using Ni-NTA Superflow (100 ml) to perform Protein tag Purification of His-tagged proteins

Get tips on using RGS·His Antibody, BSA-free (100ug) to perform Protein tag Detection of His-tagged proteins

Get tips on using Penta·His Alexa Fluor 488 Conjugate to perform Protein tag Detection of His-tagged proteins

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using DNA Isolation Kit for Cells and Tissues to perform DNA isolation / purification Cells - Primary cells Human primary keratinocytes

Get tips on using Ni-NTA Fast Start Kit (6) to perform Protein tag Purification of His-tagged proteins

Get tips on using Penta·His Antibody, BSA-free (100 ug) to perform Protein tag Detection of His-tagged proteins

Get tips on using Tetra·His Antibody, BSA-free (100 µg) to perform Protein tag Detection of His-tagged proteins

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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.