Flow cytometry protocol for thp-1 cell

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 ... 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 ...

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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 … WebOur flow cytometry protocols cover matters like sample prep of mouse and rat leucocytes, indirect staining of mononuclear total, also reducer nonspecific paint with Fc Block. Skip for main content Miss go navigation. Order Lookup. … chinese food delaware https://heating-plus.com

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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 … WebJun 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 … Web1X10^6 THP-1 cells untreated (dashed lines) or treated with LPS (red) were intracellularly stained with 0.4 ug CoraLite®488 Anti-Human IL-1 beta (CL488-66737, Clone:2A1B4) (red), or 0.4 ug Control Antibody (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C). chinese food del city

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Flow cytometry protocol for thp-1 cell

Frontiers A THP-1 Cell Line-Based Exploration of Immune …

WebPublished protocols using the promonocytic cell line THP-1 have tended to result in cells that closely resemble classically-activated macrophages, differentiated in IFNγ and LPS. … WebJan 13, 2024 · Cell Staining and Flow Cytometry Analysis. THP-1 derived M0 and iDC were cultivated in Corning® 96 Well TC-Treated Microplates (Life Sciences, Tewksbury, Massachusetts, USA) with a volume of 100 μL per well using the same protocol in the previous section on THP-1 cell Culture and Differentiation.

Flow cytometry protocol for thp-1 cell

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WebProtocol for high-throughput compound screening using flow cytometry in THP-1 cells. Flow cytometry is a valuable method for analyzing protein expressions at the single cell level but can be ... WebPlease refer to the product webpage and product-specific protocol to determine whether it is compatible with live cell staining. Collect cells by centrifugation and aspirate supernatant. Resuspend cells in 0.5–1 ml 1X PBS. Add formaldehyde to obtain a final concentration of 4%. Fix for 15 min at room temperature.

WebJul 14, 2024 · Flow cytometry analysis of treated THP-1 cells was performed for CD68, CD86, and CD206 markers to evaluate M0, M1, and M2 macrophages polarization, respectively. AGS cells were assessed for apoptosis and necrosis by flow cytometry after labeling with Annexin V-FITC and propidium iodide. 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 …

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 … WebSep 25, 2009 · THP-1 cell surface markers for Flow Cytometry - Need a robust marker for undifferentiated THP-1 cells. I am working with THP-1 cells (ATCC TIB 202), a human monocytic cell line. The majority of people that use this cell line are interested in the differentiated macrophage phenotype, which can be achieved by stimulating THP-1 cells …

WebFlow cytometry (FACS) tint protocol (Cell surface staining) Harvest, wash the cells (single fuel suspension) and adjust cell number to a concentration out 1-5x106 cells/ml in ice cold FACS Buffer (PBS, 0.5-1% BSA or 5-10% FBS, 0.1% NaN3 sodium azide*). ... same day. For extended recording (16 hr) in well when for greater flexural in planning ...

WebJan 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 … grandin road ez bed reviewsWebFeb 22, 2024 · In terms of phenotypic activation markers, measured by flow cytometry, M1 response was similar but the established MDM M2 marker CD163 was undetectable in THP-1 cells. In a beads-based assay, MDM activation did not induce significant changes, whereas M2 activation of THP-1 decreased phagocytic activity compared to M0 and M1. grandin road enter if you dareWebCell cycle assay protocols for flow cytometry. Vybrant DyeCycle Violet Stain. Vybrant DyeCycle Green and Orange Stains. Vybrant DyeCycle Ruby Stain. FxCycle Violet … grandin road decorative pillowsWebJan 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 human monocytes or monocyte-derived macrophages (MDM). ... Flow cytometry was performed to estimate the number of latex beads ingested per cell with each peak assumed to equal … chinese food del city okWebMar 11, 2024 · We describe and validate a flow cytometry-based protocol for quantifying G4 levels by using the G4-specific antibody BG4 to label standard cultured cells (Hela and THP-1) as well as primary cells obtained from human blood (peripheral blood mononuclear cells (PBMCs)). We additionally determined changes in G4 levels during the cell cycle in ... chinese food delivered in sparks nvWeb1 day ago · An increase in exposure of β-(1,3)-glucan and chitin on the cell surface was also observed through flow cytometry. Interestingly, we observed a decrease in the phagocytosis rate when the Δcdc50 mutant was co-incubated with THP-1 macrophages. The Δcdc50 mutant also exhibited weakened virulence in nematode survival tests. grandin road free shipping couponsWebSep 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. chinese food delhi