A standard angiogenic assay involves the autonomous endothelial cell response of self-organization into microvessels, also known as tubes when seeded on a basement membrane matrix in the presence of the appropriate growth factors. However, the component of basement membrane matrix may also affect the tube formation by endothelial cells. Hence it is important to use a standard angiogenesis assay kit or use the same membrane matrix with known composition to standardize the assay conditions.
A standard angiogenic assay involves the autonomous endothelial cell response of self-organization into microvessels, also known as tubes when seeded on a basement membrane matrix in the presence of the appropriate growth factors. However, the component of basement membrane matrix may also affect the tube formation by endothelial cells. Hence it is important to use a standard angiogenesis assay kit or use the same membrane matrix with known composition to standardize the assay conditions.
A standard angiogenic assay involves the autonomous endothelial cell response of self-organization into microvessels, also known as tubes when seeded on a basement membrane matrix in the presence of the appropriate growth factors. However, the component of basement membrane matrix may also affect the tube formation by endothelial cells. Hence it is important to use a standard angiogenesis assay kit or use the same membrane matrix with known composition to standardize the assay conditions.
A standard angiogenic assay involves the autonomous endothelial cell response of self-organization into microvessels, also known as tubes when seeded on a basement membrane matrix in the presence of the appropriate growth factors. However, the component of basement membrane matrix may also affect the tube formation by endothelial cells. Hence it is important to use a standard angiogenesis assay kit or use the same membrane matrix with known composition to standardize the assay conditions.
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.
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.
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.
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.
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.
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