Facs calibur s

Manufactured by BD

Sourced in United States

The BD FACS Calibur S is a flow cytometry instrument designed for cell analysis and sorting. It utilizes laser-based technology to provide high-speed, multi-parameter analysis of particles or cells suspended in a fluid stream.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions) Trypsin edta solution, by Thermo Fisher Scientific (1 mentions) Sodium azide, by Merck Group (1 mentions)

Spelling variants of this product

Calibur (250 citations) FACS Calibur-S System (8 citations) BD FACSTM (2 citations)

4 protocols using facs calibur s

CRISPR-Cas9 Genome Editing in U2OS Cells

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The assay was conducted as described^{48 (link)}. U2OS cells were seeded in 6-well plates at 2 × 10⁵ cells prior to transfection with 2 μl siRNA (20 μM) and 5 μl Lipofectamine™ 2000 (Invitrogen). One day after siRNA transfection, cells were co-transfected with 1.6 μg sgRNA plasmid pX330-LMNA1 (from Graham Dellaire) and 0.4 μg donor template pCR2.1-CloverLamin (from Graham Dellaire) and 5 μl Lipofectamine™ 2000 (Invitrogen). Gene targeting efficiency was determined by counting the percent of Clover-positive cells using a BD FACS Calibur S at 72 h post plasmids co-transfection. The results were derived from 3 to 5 transfections of at least 3 independent experiments.

Zhao W., Steinfeld J.B., Liang F., Chen X., Maranon D.G., Ma C.J., Kwon Y., Rao T., Wang W., Chen S., Song X., Deng Y., Jimenez-Sainz J., Lu L., Jensen R.B., Xiong Y., Kupfer G.M., Wiese C., Greene E.C, & Sung P. (2017). Promotion of RAD51-mediated homologous DNA pairing by BRCA1-BARD1. Nature, 550(7676), 360-365.

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Measuring Homologous Recombination Efficiency

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The DR-U2OS cell line containing a single integrated copy of the DR-GFP reporter was used^{46 (link),47 (link)}. Exponentially growing cells were seeded in 6-well plates at 2 × 10⁵ cells per well prior to transfection with 2 μl siRNA (20 μM) and 5 μl Lipofectamine™ 2000 (Invitrogen). One day after siRNA transfection, cells were transfected with 2 μg I-SceI expression vector (pCBASce) and 5 μl Lipofectamine™ 2000. HR proficiency was determined by counting the fraction of GFP-positive cells using a BD FACS Calibur S at 72 h post I-SceI transfection. The results were derived from 3 to 5 transfections of at least 3 independent experiments.

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Apoptosis Analysis of JK184 Treatment

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Cells were plated at 5 × 10⁴ cells/well in 6-well format and allowed to adhere overnight. The following day, cells were treated with 0.02 μM JK184 or 0.0002% DMSO vehicle control. After four days, floating cells were combined with adherent cells harvested by trypsinization and analyzed by flow cytometry using the BD Biosciences Pharmingen (San Diego, CA, USA) FITC Annexin V Apoptosis Detection Kit I. Samples were analyzed with the BD FACScaliburS flow cytometer with recording of 15,000 events per sample. Each line was treated independently and analyzed in three biological replicates. Gates were based on negative control signals, and plots generated using FlowJo 8.8.2 (Tree Star, Inc., San Carlos, CA, USA).

Colavito S.A., Zou M.R., Yan Q., Nguyen D.X, & Stern D.F. (2014). Significance of glioma-associated oncogene homolog 1 (GLI1)expression in claudin-low breast cancer and crosstalk with the nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) pathway. Breast Cancer Research : BCR, 16, 444.

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Characterization of Isolated MSC Surface Markers

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Cell surface markers on isolated MSCs at passage 8 to 10 were determined by flow cytometry on BD FACSCaliburs (BD Biosciences). Briefly, cells were lifted by 0.05% Trypsin-EDTA solution (Invitrogen) and pelleted at 1000 g for 3 minutes, washed thrice with 1x PBS supplemented with 2% FBS (Hyclone) and 10 mM sodium azide (Sigma-Aldrich, St. Louis, MO, USA), incubated with antibody conjugated with FITC for 20 minutes on ice. After antibody incubation, cells were pelleted at 1000 g for 3 minutes and washed thrice with 1x PBS supplemented with 2% FBS (Hyclone) and 10 mM sodium azide (Sigma-Aldrich). Cells were kept in 1x PBS supplemented with 2% FBS (Hyclone) and 5 mM sodium azide (Sigma-Aldrich) before analysis at 4 °C. Antibodies against cell surface markers were: anti-CD11b-FITC, anti-CD45-FITC, anti-Sca-1-FITC, anti-CD29-FITC; proper IgG isotype controls were used to assess background signal. All antibodies were purchased from BioLegend (San Diego, CA, USA) and used at the dilution ratios recommended by manufacturer.

Wu H., Gordon J.A., Whitfield T.W., Tai P.W., van Wijnen A.J., Stein J.L., Stein G.S, & Lian J.B. (2017). Chromatin Dynamics Regulate Mesenchymal Stem Cell Lineage Specification and Differentiation to Osteogenesis. Biochimica et biophysica acta, 1860(4), 438-449.

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