CRISPR Rat Deletion

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.

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 Alexa Fluor® 647 Rat Anti-Mouse CD206 to perform Flow cytometry Anti-bodies Mouse - CD206

Get tips on using PE-Cy™7 Rat Anti-Mouse CD86 to perform Flow cytometry Anti-bodies Mouse - CD86

Get tips on using Alexa Fluor® 700 Rat Anti-Mouse CD86 to perform Flow cytometry Anti-bodies Mouse - CD86

Get tips on using PerCP-Cy™5.5 Rat Anti-Mouse CD19 to perform Flow cytometry Anti-bodies Mouse - CD19

Get tips on using PerCP-Cy™5.5 Rat Anti-Mouse CD8a to perform Flow cytometry Anti-bodies Mouse - CD8a

Get tips on using PerCP-Cy™5.5 Rat Anti-Mouse CD4 to perform Flow cytometry Anti-bodies Mouse - CD4

Get tips on using Alexa Fluor® 647 Rat anti-Mouse CD34 to perform Flow cytometry Anti-bodies Mouse - CD34

Isolating RNA from tissues and paraffin-embedded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the integrity of RNA