Staining buffer

Manufactured by R&D Systems

Staining buffer is a laboratory reagent used to prepare samples for staining procedures. It is formulated to maintain the integrity and structure of cells or tissues during the staining process. The buffer composition helps to optimize staining conditions and ensure consistent and reliable staining results.

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Lab products found in correlation

Intracellular fixation and permeabilization kit, by BD (1 mentions) Cd206 pe, by Thermo Fisher Scientific (1 mentions) F4 80 fitc, by Thermo Fisher Scientific (1 mentions) Facsaria flow cytometer, by BD (1 mentions) Cd16 32, by BD (1 mentions) Inos apc, by Thermo Fisher Scientific (1 mentions) Anti cd14 pe, by Thermo Fisher Scientific (1 mentions) Anti mouse cd16 cd32, by BD (1 mentions) Accuri flow cytometer, by BD (1 mentions) Facscalibur flow cytometer, by BD (1 mentions)

Spelling variants of this product

Flow cytometry staining buffer (22 citations) Flow Cytometry Staining Buffer (1X) (3 citations)

3 protocols using staining buffer

Macrophage Phenotyping by Flow Cytometry

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Samples were incubated with an Fc receptor blocker (CD16/32, BD Bioscience) to reduce nonspecific antibody binding. Freshly isolated samples were resuspended in staining buffer (R&D Systems) and stained with F4/80‐FITC (eBioscience) for 30 min at 4 °C. For intracellular staining, we used the Intracellular Fixation and Permeabilization Kit (BD Bioscience); this was used in accordance with the manufacturer’s instructions. Cells were then washed and stained with iNOS-APC (eBioscience) and CD206-PE (eBioscience). Flow cytometry was performed using a FACS Aria flow cytometer (BD Bioscience) and data were analyzed with FlowJo V10.6.2 software (TreeStar, Ashland, OR).

Zhang Y., Cai Z., Shen Y., Lu Q., Gao W., Zhong X., Yao K., Yuan J, & Liu H. (2021). Hydrogel-load exosomes derived from dendritic cells improve cardiac function via Treg cells and the polarization of macrophages following myocardial infarction. Journal of Nanobiotechnology, 19, 271.

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Flow Cytometric Analysis of LPS-Cell Interactions

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Flow cytometry was used to detect cell surface receptors and fluorescein isothiocyanate-conjugated LPS (FITC-LPS) associated with intact cells as previously described with minor modifications [40 (link)]. For the detection of cell surface receptors, cells were pretreated with fatty acid as indicated above with or without ultra-pure LPS for stimulation. One million cells were blocked with 1 μg anti-mouse CD16/CD32 (BD Biosciences, San Jose, CA) in 100 μL for 5 min at 4°C and then labeled with 0.25 μg anti-TLR4-APC (R&D Systems), 0.5 μg anti-TLR4/MD2-APC (eBioscience), 0.5 μg anti-CD14-PE (eBioscience), or their isotype controls in 100 μL blocking solution for 30 min at room temperature. To assess the effect of LPS-cell association, fatty acid-treated cells were harvested and suspended in the original culture media containing the treatment fatty acid, and incubated with LPS-FITC (1 μg/mL final concentration) for 1 h at 37°C. Fluorescent labeled cells were washed and resuspended in the staining buffer (R&D Systems), and analyzed on an Accuri Flow Cytometer (BD Biosciences).

Honda K.L., Lamon-Fava S., Matthan N.R., Wu D, & Lichtenstein A.H. (2014). EPA and DHA exposure alters the inflammatory response but not the surface expression of toll-like receptor 4 in macrophages. Lipids, 50(2), 121-129.

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Multilineage Differentiation of Isolated MSCs

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Multilineage differentiation potential of isolated MSCs was confirmed by subculturing MSCs on chamber slides and inducing differentiation to adipogenic, chondrogenic, and osteogenic cell lineages using specific conditioning media. Histochemical analysis was performed by oil red O, alizarin red S, and alcian blue as previously described.²⁰ In addition, ADSC, BMSC, and DPSC were characterized to confirm their stemness profile before endothelial induction. For this, expression of typical MSCs markers was evaluated by flow cytometry using a Human MSC Analysis BD Stemflow™ kit (BD Biosciences). In brief, 5 × 10⁵ cells corresponding to ADSC, BMSC, and DPSC were placed in flow cytometry tubes and washed with 2 ml of staining buffer (R&D Systems Inc). Then, Fc receptors were blocked by incubating the cells for 5 min with 2 ml of PBS containing 0.1% bovine serum albumin and 0.1% FBS. Next, cells were stained with a positive cocktail (FITC CD90, PerCP‐Cy CD105, and APC CD73) and a negative MSC cocktail (PE CD45, PE CD34, PE CD11b, PE CD19, and PE HLA‐DR) and incubated for 45 min at 4ºC in darkness. Once cells were labeled, cells were placed in a staining buffer and analyzed using a FACSCalibur flow cytometer (BD Biosciences).

Blanco‐Elices C., Chato‐Astrain J., Oyonarte S., Bermejo‐Casares F., España‐López A., Fernández‐Valadés R., Sánchez‐Quevedo M.D., Alaminos M., Martín‐Piedra M.A, & Garzón I. (2021). Generation of a novel model of bioengineered human oral mucosa with increased vascularization potential. Journal of Periodontal Research, 56(6), 1116-1131.

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