Flow cytometry protocol for thp-1 cell

WebFor the detection of the viability of Jurkat and THP-1 cells, an MTT assay was employed as previously described . Briefly, both Jurkat and THP-1 cells in the logarithmic growth phase were seeded at a density of 1 × 10 5 cells/well on a 24-well plate and treated with various concentrations (40, 20, 10, 5 and 2.5 µM) of each compound. After the ... WebFlow cytometry agreement & procedures including; straightforward staining, directly staining of intracellularly antigen & cytokines, single preparation & permeabilization. 425805 392e8725-8a05-419c-a0a8-99ec1752bb34

Protocol for high-throughput compound screening using flow cytometry in ...

WebSep 1, 2015 · THP-1, a human monocytic cell line and its derived macrophages serve as a useful cell culture model for mechanistic studies of hCMV infection and its underlying biology. A major methodological challenge is the lack of a quick and reliable tool to accurately determine the efficiency of hCMV infection in THP-1 derived macrophages. WebTo get M1-macrophages: 24-48h of 50ng/ml IFNg and 50 ng/ml LPS. To get M2-macrophages: 24-48h of IL-4 and IL-13 (25 ng/ml both). In summary, when I go to flow … bisson ford padova https://ajliebel.com

Quantitative analysis of cell proliferation by a dye dilution assay ...

WebJan 21, 2024 · Affiliation Flow Cytometry Laboratory, Botucatu Medical School - UNESP, Botucatu, Brazil ⨯ . ... under protocol 2.018.399. THP-1 culture and differentiation. THP-1 cells were cultured in 25 cm 2 sterile … WebCell cycle assay protocols for flow cytometry. Vybrant DyeCycle Violet Stain. Vybrant DyeCycle Green and Orange Stains. Vybrant DyeCycle Ruby Stain. FxCycle Violet … WebJun 5, 2015 · At 24 hpi, IE2-EYFP positive cells were measured by flow cytometry; (b) THP-1 monocytes were differentiated to THP-1 derived macrophages by the addition of PMA for 48 h followed by infection of the obtained differentiated THP-1 cells with TB40E/IE2-EYFP at an MOI of 0.1. At 24 hpi, IE2-positive cells were detected by flow … bisson innovations

Quantitative analysis of cell proliferation by a dye dilution assay ...

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Flow cytometry protocol for thp-1 cell

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WebPrepare your cell suspensions for Flow Cytometry. Although most flow cytometry experiments involve labeling populations of cells that are relatively abundant, the number of cells required will vary depending upon the rarity of your cells. In general, researchers will stain between 1 x 10 5 and 1 x 10 6 cells per sample. WebJun 1, 2024 · This protocol provides instructions to perform a high-throughput small molecule screen using flow cytometry analysis of THP-1 cells, a human monocytic leukemia cell line. We describe a …

Flow cytometry protocol for thp-1 cell

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WebMar 9, 2015 · 1.3 For suspension cells, centrifuge to obtain a cell pellet and aspirate the supernatant. Resuspend the cells gently in prewarmed (37°C) probe-containing medium. Incubate the cells for 30 minutes to 2 hours under growth conditions appropriate for the particular cell type (see note above regarding internalization rate of these probes). Re ... WebJun 26, 2024 · We found that the baseline median fluorescent intensity (MFI) of THP-1 cells changed depending on the time of sample acquisition to the flow cytometer. To solve this problem, a fixation step was introduced in different stages of the assay's protocol, obtaining more reproducible and sensitive results when a post-LC3 staining fixation was ...

WebMar 18, 2024 · This protocol provides instructions to perform a high-throughput small molecule screen using flow cytometry analysis of THP-1 cells, a human monocytic … WebApr 14, 2024 · A similar approach has been used to characterize the adherence and transmigration properties of peripheral blood mononuclear cells on human aortic endothelial cells and brain microvascular endothelial cells . One limitation of the flow cytometry-based assay was the inherent variability in the cells used; for example, the passage number of …

WebOct 1, 2024 · Cell yields resulting from THP-1 differentiation in plates (pDMs) and differentiation in tubes (tDMs) were compared by flow cytometry. Cells were … WebFlow Cytometry Reagents. Clinical Diagnostics; ... Single-color flow cytometric analysis of Ig Isotype control staining on THP-1 cells. THP-1 cells were incubated at room …

Web5 Collect 2 million cytotoxic T cells per sample (i.e. per well) from an on-going culture 6 Spin them down at 350 x gfor 00:05:00 at 4 °C and re-suspend them in fresh media at 1 million per mL concentration 7 Supplement T cells with 200 IU/mL rIL2 8 Aspirate the culture media from each of the 24-well-plate wells that contain a sample. Try to aspirate as much as …

WebFeb 3, 2024 · Single-cell preparation from different organs, staining of cells with fluorophore-conjugated antibodies, and analysis of the stained cells using flow … bisson houlgateWebFeb 3, 2024 · Single-cell preparation from different organs, staining of cells with fluorophore-conjugated antibodies, and analysis of the stained cells using flow cytometry are as described 51. Briefly, cells ... darth ruynWebJan 12, 2024 · Once the cells are suspended, seal the tube, invert a few times, and then divide the cells into the freezing vials. Transfer the vials to your equipment for freezing. Do Not Leave Your THP-1 Cells in the … darths and droids tv tropesWebFlow Cytometry Blocking Controls Fc receptors are found on monocytes, macrophages, dendritic cells and B cells. ... have been developed which, when added to a staining protocol, can ensure that only antigen specific … darth sand liar im breaking your neckWebThe first step to isolating your cells of interest begins with forward scatter (FSC) and side scatter (SSC). Larger, more complex cells will be higher in both parameters. Knowing the size and makeup of your cells of interest is key to gating accurately. If cell lines are being used, the FSC/SSC should show one main population of cells: this ... darths and droids hosnianWebJan 13, 2010 · We have compared the phenotype of the promonocytic THP-1 cell line after various protocols of differentiation utilising VD3 and PMA in comparison to primary … darthsandr downloadbisson innovations inc