ChIP H3K36Me3 Bovine Rat

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Get tips on using ON-TARGETplus Human RET (5979) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - TT RET

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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.

Cellular assays Live / Dead assay mammalian cells bmMSCs human bone marrow

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.

Cellular assays Live / Dead assay mammalian cells mouse bone marrow-derived macrophages

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 U2OS (human bone osteosarcoma epithelial cells)

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - Mouse Bone

Products Thermo Fisher Scientific TRIzol Reagent

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - Human Bone

Products Thermo Fisher Scientific TRIzol Reagent

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - mouse bone tissue

Products Thermo Fisher Scientific TRIzol Reagent

Get tips on using miRNeasy Mini kit to perform RNA isolation / purification Tissue - Mouse Bone

Products Qiagen miRNeasy Mini kit

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - Mouse Bone

Products Qiagen RNeasy Mini Kit

Get tips on using RNeasy Micro Kit to perform RNA isolation / purification Tissue - Human Bone

Products Qiagen RNeasy Micro Kit

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