Flojo

Manufactured by Tree Star

Sourced in United States

FloJo is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative-scale separations. It features precise flow control, advanced detector options, and user-friendly software for efficient and reliable chromatographic analysis.

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Lab products found in correlation

Facscalibur, by BD (7 mentions) Lsr 2 flow cytometer, by BD (3 mentions) Attune flow cytometer, by Thermo Fisher Scientific (3 mentions) Cd64 x54 5 7, by BioLegend (3 mentions) Cd11b m1 70, by BD (3 mentions) Cd11c hl3, by BD (3 mentions) I a 1 e m5 114, by BioLegend (3 mentions) F4 80 bm8, by Thermo Fisher Scientific (3 mentions) Viability 65 0863 14, by Thermo Fisher Scientific (3 mentions) Facsaria 2 flow cytometer, by BD (3 mentions) Facsaria, by BD (3 mentions) Truestain, by BioLegend (2 mentions) Lsrfortessa flow cytometer, by BD (2 mentions) High throughput sampler, by BD (2 mentions) Lsrfortessa cell analyzer, by BD (2 mentions) Cd4 rm4 5, by BD (2 mentions) Ebio13a, by Thermo Fisher Scientific (2 mentions) Tc11 18h10, by BD (2 mentions) Ly6g 1a8, by BioLegend (2 mentions) Ly6c hk1, by BioLegend (2 mentions)

38 protocols using flojo

Characterization of Endothelial Progenitor Cells

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The peripheral blood mononuclear cells were suspended in saline and incubated with fluorescein isothiocyanate anti-mouse stem cells antigen (Sca)-1 (Invitrogen, 11-5981-82, Carlsbad, CA, USA) and phycoerythrin anti-mouse vascular endothelial growth factor receptor 2, also known as Flk-1 (Invitrogen, 12-5821-82, Carlsbad, CA, USA) at room temperature for 30 min. A BD FACScalibur flow cytometer (BD, East Rutherford, NJ) was used, and data were analyzed with FloJo (Treestar). Data are presented as % gated, relative to control group.
EPCs were harvested and washed prior to suspension in phosphate-buffered saline (PBS) for flow cytometry. To identify the characteristics of EPCs, VE-cadherin (Biolegend, 348505, San Diego, CA), CD31 (Biolegend, 303117, San Diego, CA), CD34 (BD, 555821, East Rutherford, NJ), KDR (R&D, FAP357, Minneapolis, MN, USA), CD133 (MACS, 130-111-756, Germany), CD3 (Biolegend, 300407, San Diego, CA), CD68 (Biolegend, 333805, San Diego, CA), CD86 (Biolegend, 374203, San Diego, CA), CD163 (Biolegend, 33605, San Diego, CA), and CD206 (Biolegend, 321123, San Diego, CA) antibodies were used. Cells were analyzed by BD FACScalibur flow cytometer (BD, East Rutherford, NJ), and data were analyzed with FloJo (Treestar). Data are presented as % gated.

Chen C., Lin L.Y., Chen J.W, & Chang T.T. (2023). CXCL5 suppression recovers neovascularization and accelerates wound healing in diabetes mellitus. Cardiovascular Diabetology, 22, 172.

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Cell Cycle Analysis of XAV939 Treatment

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Cells were grown for 24 h in complete DMEM and then incubated with free and conjugated XAV939 dissolved in fresh DMEM for 48 h. After which, cells were harvested using trypsin, washed with phosphate buffered saline (PBS), fixed in ice-cold ethanol (70%) and kept at –20°C. Fixed cell suspensions were centrifuged at 1,500 rpm for 7 min and the cell pellet was redispersed in PBS. Cells were treated with 200 μg/mL RNAse (Sigma) for 30 min at 37°C. Following which, DNA staining with 100 μg/mL of propidium iodine (Sigma) was performed at room temperature for 15 min. A BD LSR II flow cytometer (BD Biosciences) with 488 nm excitation laser and fluorescence detection in the PE channel was used to measure the cell cycle distribution (15,000 events were acquired for each sample). The obtained data was analyzed using FloJo (Tree Star Inc.), a flow cytometry analyzing software. The amount of propidium iodide intercalated to DNA was used as a parameter to determine the cell cycle distribution phases.

Afifi M.M., Austin L.A., Mackey M.A, & El-Sayed M.A. (2014). XAV939: From a small inhibitor to a potent drug bioconjugate when delivered by gold nanoparticles. Bioconjugate chemistry, 25(2), 207-215.

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Flow Cytometry Analysis of SMIM1 Variants

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Flow cytometry analysis was performed as described previously.5 HEK293T cells were transfected with either empty pcDNA3.1+ vector (Invitrogen), wild‐type SMIM1, or SMIM1*c.161T>C (double‐stranded DNA ordered at gBlocks IDT) as described previously.5 Antibodies used were as follows: immunopurified anti‐Vel (MB46) as primary antibody and Alexa Fluor‐647 anti‐human IgG (Invitrogen) as secondary antibody. Results were acquired on a flow cytometer (BD CantoII/LSRII/Fortessa, BD Bioscience, Breda, the Netherlands) and analyzed using computer software (FloJo, Treestar).

van der Rijst M.V., Voorn L., Veldhuisen B., Jongerius J.M., van den Akker E, & van der Schoot C.E. (2019). Identification of a novel single‐nucleotide mutation in SMIM1 gene that results in low Vel antigen expression. Transfusion, 59(10), E8-E10.

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Analyzing Polyfunctional Cytokine Production

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We used the paired Student’s t test to compare ORF59 PF-8^pos and ORF59 PF-8^neg cells within groups and an ANOVA for comparisons between groups. Flow cytometry data were analyzed using FloJo software (Tree Star). Polyfunctional cytokine and chemokine production levels were assessed by using SPICE permutation tests and Student’s t test (version 4.3; M. Roederer, Vaccine Research Center, NIAID, NIH).

Knowlton E.R., Rappocciolo G., Piazza P., Lepone L.M., Nadgir S.V., Bullotta A., Berendam S.J., Li J., Reinhart T.A., Jenkins F.J, & Rinaldo C.R. (2014). Human Herpesvirus 8 Induces Polyfunctional B Lymphocytes That Drive Kaposi’s Sarcoma. mBio, 5(5), e01277-14.

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Murine Lung Cell Phenotyping

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Single cell suspensions were produced from murine lung (n≥5) after irradiation. After blocking the Fc receptor with anti-mouse CD16/CD32 antibody (101335), cells were labeled with fluorophore-conjugated BioLegend antibodies against mouse CD11b (101217), CD206 (141726), and F4/80 (123116). Labeled cells were fixed with 2 % PFA and acquired 100,000 events for each sample with LSR Fortessa (BD Biosciences, Franklin Lakes, NJ) and analyzed with FloJo (Tree Star, Inc., Ashland, OR).

Chung E.J., Kwon S., Reedy J.L., White A.O., Song J.S., Hwang I., Chung J.Y., Ylaya K., Hewitt S.M, & Citrin D.E. (2020). IGF-1 receptor signaling regulates type II pneumocyte senescence and resulting macrophage polarization in lung fibrosis.. International journal of radiation oncology, biology, physics, 110(2), 526-538.

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Imaging-based Analysis of sfGFP

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TZM-bl cells were plated in glass-bottom 96-well optical imagining microplates (Brooks Life Science, Chelmsford, MA) and incubated at 37°C O/N. Cells were exposed to constructs or control media in the presence or absence of the indicated inhibitors or controls for 1 h at 37°C. Cells were stained with 0.1 μg/mL Hoechst for 30 min, washed, and phenol red free media supplemented with 10 mM HEPES was added. Cells were then imaged with the iCys Research Imaging Cytometer (CompuCyte, Thorlabs Inc., Newton, NJ) using a 60X (Figure 4) or 40X (Supplementary Fig. 3) 0.9 NA objective. sfGFP was excited using a laser at 488 nm and fluorescence was collected with a 515–545 nm emission filter. Ten fields were imaged with biological triplicates for each condition specified. Using the iCyte (CompuCyte) imaging software individual cells were identified and analyzed for green fluorescence. Data were exported to FloJo (Treestar, Ashland, OR) for analysis of cytometric data.

Costantini L.M., Irvin S.C., Kennedy S.C., Guo F., Goldstein H., Herold B.C, & Snapp E.L. (2014). Engineering and exploitation of a fluorescent HIV-1 gp120 for live cell CD4 binding assays. Virology, 476, 240-248.

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Kupffer Cell and MDSC Characterization

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Cells were incubated with Fc-blocking reagent (TrueStain, Biolegend) for 5 min, and subsequently stained for 20 min at 4°C with the relevant antibody. Some antibodies were used to analyze Kupffer cells: FITC anti-F4-80 (BM8), APC anti-I-A-I-E (M5/114.15.2), PE anti-CD86 (GL-1), PECy7 anti-CD68 (FA-11), PerCP anti-CD11b (M1/70) (Biolegend), PE anti-CD273 (122), and anti-CD274 (MIH5) (eBiosciences). For experiments involving Kupffer cell/MDSC co-culture, Kupffer cells were first labeled with CFSE (0.5 µM, Invitrogen). MDSC were characterized with the following antibodies: APC anti-CCR2 (475301, R&D Systems), APC anti-CD244.2 (eBio244F4, eBiosciences), APC anti-CD54 (YN1/1.7.4), APC anti-CD115 (AFS98), PE anti-Ly6C (HK1,4), PerCP anti-CD11b (M1/70), AlexaFluor488 anti-CD146 (ME-9F1), PECy7 anti-Gr1 (RB6-8C5), APC Cy7 anti-CD45 (30-F11, Biolegend). All samples were acquired with a Flow Cytometer Cyan (Beckman Coulter), and analyzed with FloJo (Treestar).

Lacotte S., Slits F., Orci L.A., Meyer J., Oldani G., Delaune V., Gonelle-Gispert C., Morel P, & Toso C. (2016). Impact of myeloid-derived suppressor cell on Kupffer cells from mouse livers with hepatocellular carcinoma. Oncoimmunology, 5(11), e1234565.

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ROS Quantification in Viable Cells

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Four days after transduction with TAZ or control shRNA sequences cells were stained with Carbocy-H2DCFDA in PBS buffer at 37°C for 30 minutes, and resuspended in PBS with the viability dye PI to assess the reactive oxygen species produced by viable cells. Data was acquired by LSRFortessaX20 (BD Biosciences, FL, USA), and MFI of Carbocy-H2DCFDA in PI negative cells were analyzed using FloJo (TreeStar, OR, USA).

Seneviratne A.K., Xu M., Aristizabal Henao J.J., Fajardo V.A., Hao Z., Voisin V., Xu G.W., Hurren R., Kim S., MacLean N., Wang X., Gronda M., Jeyaraju D., Jitkova Y., Ketela T., Mullokandov M., Sharon D., Thomas G., Chouinard-Watkins R., Hawley J.R., Schafer C., Yau H.L., Khuchua Z., Aman A., Al-awar R., Gross A., Claypool S.M., Bazinet R.P., Lupien M., Chan S., De Carvalho D.D., Minden M.D., Bader G.D., Stark K.D., LeBlanc P, & Schimmer A.D. (2019). The Mitochondrial Transacylase, Tafazzin, Regulates AML Stemness by Modulating Intracellular Levels of Phospholipids. Cell stem cell, 24(4), 621-636.e16.

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Quantifying UPR under Ethanol Stress

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To measure UPR activity under ethanol stress we used a modified plasmid reporter called UPR-mCherry that encodes red fluorescent protein mCherry (Merksamer et al., 2008 (link)), driven by a minimal CYC1 promoter and four tandem unfolded protein response elements. Cells containing the plasmid were grown overnight in GPY with geneticine medium at 28°C and were allowed to reach the early exponential phase (an approximate OD600 value of 0.4) for the analysis. Then the culture was divided into sterile centrifuge tubes, pelleted and incubated with GPY media with geneticine, with or without 10% (v/v) ethanol. Cells were grown at 28°C, sampled every 2 h, pelleted and frozen in liquid nitrogen until use. GFP fluorescence was measured by flow cytometry in a LSR Fortessa flow cytometer (BD Biosciences) and analyzed with the FACS DIVA software to compile.fcs files. Files were analyzed using FloJo (Tree Star Ashland, OR). Median fluorescence intensities (MFI) were calculated for each channel and were normalized with time cero sample data. To quantify UPR induction, fractional area (fa) of 10% ethanol condition was normalized against control (see above). Biological triplicates were performed in all cases.

Navarro-Tapia E., Nana R.K., Querol A, & Pérez-Torrado R. (2016). Ethanol Cellular Defense Induce Unfolded Protein Response in Yeast. Frontiers in Microbiology, 7, 189.

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GFP and mCherry Degradation Dynamics

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Data for GFP and mCherry degradation dynamics and propidium iodide staining were collected using an LSRFortessa cell analyzer equipped with a High Throughput Sampler (BD Biosciences), and data for the synthetic toggle switch were collected using a FACSAriaII flow cytometer (BD Biosciences). For each GFP and mCherry measurement, cells were fixed in 1% paraformaldehyde (PFA), held at 4° for up to 5 days, and then diluted 1:10 in PBS for analysis. At least 5,000 cells were collected for each measurement and FloJo (Treestar) was used for data analysis.

Cameron D.E, & Collins J.J. (2014). Tunable protein degradation in bacteria. Nature biotechnology, 32(12), 1276-1281.

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