rna-isolation-purification-cells-primary-porcine-primary-airway-epithelial-cell

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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.

Cellular assays Apoptosis assay cell type Sao2-2, MG-62

Get tips on using Ni-NTA Agarose to perform Protein expression and purification Insect cells - Hi5 TYR

Products Qiagen Ni-NTA Agarose

Get tips on using TRI Reagent® to perform Protein isolation Mammalian cells - Mouse_Brown fat

Products Sigma-Aldrich TRI Reagent®

Get tips on using In Situ Cell Death Detection Kit, TMR red to perform TUNEL assay cell type - A549, NCI-H460, H1299 human lung cancer cells

Products Sigma-Aldrich In Situ Cell Death Detection Kit, TMR red

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 MT-2 (human T cell leukaemia)

Get tips on using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation to perform Live / Dead assay mammalian cells - Spleen cells

Products Thermo Fisher Scientific LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit, for UV excitation

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - 3T3-L1

Products Sigma-Aldrich RIPA Buffer

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - THP-1

Products Sigma-Aldrich RIPA Buffer

Get tips on using Trichloroacetic acid to perform Protein isolation Mammalian cells - THP-1

Products Sigma-Aldrich Trichloroacetic acid

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - CHO-K1

Products Sigma-Aldrich RIPA Buffer

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