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Pe texas red

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PE Texas Red is a fluorescent dye used in molecular biology and cell biology applications. It has an excitation maximum at 596 nm and an emission maximum at 615 nm, making it suitable for detection in the red region of the visible spectrum.

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Lab products found in correlation

Pe cy7, by BioLegend (3 mentions) Permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Ber act8, by BioLegend (2 mentions) Brilliant violet 605, by BioLegend (2 mentions) Alexa fluor 647, by BioLegend (2 mentions) Fixation buffer, by Thermo Fisher Scientific (2 mentions) Brilliant violet 421, by BioLegend (2 mentions) Buv395, by BD (2 mentions) Brilliant violet 650, by BioLegend (2 mentions) Sj25 c1, by Thermo Fisher Scientific (2 mentions) Hi100, by BioLegend (2 mentions) Percpfl710, by Thermo Fisher Scientific (2 mentions) Uchl1, by Thermo Fisher Scientific (2 mentions) 6 laser lsrii analytical flow cytometer, by BD (2 mentions) Alexa fluor 488, by BioLegend (2 mentions) Fluorescein isothiocyanate (fitc), by BD (2 mentions) Apc cy7, by BD (2 mentions) Lightning link conjugation kit, by Abcam (1 mentions) Cd150, by BioLegend (1 mentions) Facsaria, by BD (1 mentions)

7 protocols using pe texas red

1

Fluorescent Labeling of Lectins

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For covalent labeling of lectins with fluorophores targeting the primary amine groups of lectins, Lightning Link conjugation kits were used following the simple and rapid procedure provided by the manufacturer (Abcam, Cambridge, US). Aleuria Aurantia lectin (AAL, Cat. L-1390-2, Vector Laboratories, Newark, USA) was labeled with APC (Cat. ab201807, Lot.GR3388854-1), recombinant human galectin-1 as a gift of Dr. Éva Monostori, Gal-1, expression and purification steps described in detail in (44 (link)–46 (link)), that was conjugated to PE/Cy7 (Cat. ab102903, Lot. GR3390407-1), recombinant human sialoadhesin, Siglec-1/CD169 protein (Siglec-1, Cat. 5197SL050, Fischer Scientific, Massachusetts, USA) was labeled with PE/Texas Red (Cat. ab269899, Lot. GR3395603-2), and finally galectin-3 (Gal-3, Cat. 450-38, PeproTech, London, UK) was chemically linked to APC/Cy7 dye (Cat. ab102859, Lot. GR3396716-1). Fluorescein-conjugated Sambucus nigra agglutinin was purchased from Vector Laboratories (Cat. FL-1301-2). The list of the selected lectins for cytometry is listed in Supplementary Table 4.
Szabó E., Faragó A., Bodor G., Gémes N., Puskás L.G., Kovács L, & Szebeni G.J. (2024). Identification of immune subsets with distinct lectin binding signatures using multi-parameter flow cytometry: correlations with disease activity in systemic lupus erythematosus. Frontiers in Immunology, 15, 1380481.
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2

Isolation and Characterization of Murine LSK Cells

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Bone marrow cells were flushed from the tibias and femurs of mice with PBS without calcium or magnesium. For detection of LSK cells, whole bone marrow cells were incubated with biotin-conjugated monoclonal antibodies to the following lineage markers: B220 (6B2), CD4 (GK1.5), CD8 (53–6.7), Gr1 (8C5), Mac1 (M1/70), Ter119 (Ter119), and interleukin-7 receptor (IL-7R) (A7R34); FITC-conjugated antibody to Sca1 (Ly6A/E; D7) and APC-conjugated antibody to c-Kit (ACK2). CD48 and CD150 were measured with PE-conjugated antibody to CD48 (HM48-1) and PE Cy7-conjugated antibody to CD150 (TC15-12F 12.2). All antibodies were purchased from eBioscience except for antibody CD150, which was purchased from Biolegend. Biotin-conjugated lineage marker antibodies were detected using streptavidin-conjugated PE Texas Red (ThermoFisher). Antibodies to B220, CD4, CD8, Gr1, Mac1 and IL-7R were diluted 1:550. Antibodies to Ter119 and Gr1 were diluted 1:275. Antibodies to Sca1, c-Kit, CD48, and CD150 were diluted 1:80. Non-viable cells were excluded using the viability dye DAPI (1 μg ml−1). Cells were sorted with a FACSAria (Becton Dickinson) automated cell sorter. Analyses were performed on a FACSCanto or LSR II flow cytometer (Becton Dickinson). Data were analyzed using FlowJo software (Tree Star).
Nguyen-McCarty M, & Klein P.S. (2017). Autophagy is a signature of a signaling network that maintains hematopoietic stem cells. PLoS ONE, 12(5), e0177054.
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3

Multiparametric Flow Cytometry Analysis

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The following flurochrome-conjugated antibodies were used for surface staining: anti-human CD3 (Brilliant Violet 650, 1:100, OKT3, BioLegend, San Diego, CA), CD4 (PeCy7, 1:100, SK3, BioLegend), CD8 (APC-Cy7, 1:100, SK1, BD Biosciences), CD19 (PE Texas Red, 1:100, SJ25-C1, Invitrogen or APC, HIB19, Biolegend), CD25 (FITC, 1:50, 2A3, BD Biosciences, San Jose, CA), CD31 (APC, 1:100, WM59, eBioscience, San Diego, CA), CD45RA (Brilliant Violet 605, 1:100, HI100, BioLegend), CD45RO (PerCpFl710, 1:100, UCHL1, eBioscience), CD69 (Brilliant Violet 421, 1:100, FN50, BioLegend or BUV395, 1:100, FN50, BD Biosciences), CD103, (Alexa Fluor 647, 1:100, Ber-ACT8, BioLegend), CD127 (BV711, 1:100, A019D5, BioLegend), CCR7 (Alexa Fluor 488, 1:100, TG8, BioLegend). For intracellular staining, surface stained cells were resuspended and incubated in fixation buffer (eBioscience), washed, resuspended in 0.1mL permeabilization buffer (eBioscience) and stained with anti-Foxp3 antibodies (PE, 1:20, 236A/E7, eBioscience) and Ki67 (α700, 1:100, Ki-67, BioLegend) for 30 min at room temperature and washed twice with permeabilization buffer. Stained cells were acquired on a 6-laser LSRII analytical flow cytometer (BD Biosciences) in the CCTI flow cytometry core and analyzed using FlowJo software (Treestar, Ashland, OR).
Thome J.J., Bickham K.L., Ohmura Y., Kubota M., Matsuoka N., Gordon C., Granot T., Griesemer A., Lerner H., Kato T, & Farber D.L. (2015). Early life compartmentalization of human T cell differentiation and regulatory function in mucosal and lymphoid tissues. Nature medicine, 22(1), 72-77.
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4

Thymus Tissue Dissociation and Flow Cytometry

Cited 2 times
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Thymus tissue was incubated for 1 h at 37°C in collagenase digestion mixture (0.1 mg/ml collagenase type 4 in calcium-replete HBSS containing 10% FCS and 200 U/ml DNase). After homogenization and filtration, the resultant cells were stained and analyzed using an LSRII flow cytometer together with FlowJo software (Tree Star) as has been described [35 (link),36 (link)]. Biotin-conjugated anti-CD90.2 (53–2.2), eFluor605NC-conjugated anti-CD19 (1D3), APC-conjugated anti-CD4 (GK1.5), eFluor450-conjugated anti-CD8 (53–6.7), and PE-Cy7-conjugated anti-CD3 (145–2C11) were purchased from eBioscience, whereas APC-Cy7-conjugated anti-CD45 (30-F11) was obtained from BD Pharmingen. Biotin-conjugated antibodies were detected using streptavidin conjugated with PE-TexasRed (Invitrogen). Mann–Whitney t-tests (with a 95% confidence interval) were performed using Prism 6.0 (GraphPad Software, La Jolla, USA). All P-values are two-tailed (*P < 0.05, **P < 0.01, and ***P < 0.001).
Chauhan S., Diril M.K., Lee J.H., Bisteau X., Manoharan V., Adhikari D., Ratnacaram C.K., Janela B., Noffke J., Ginhoux F., Coppola V., Liu K., Tessarollo L, & Kaldis P. (2016). Cdk2 catalytic activity is essential for meiotic cell division in vivo. The Biochemical journal, 473(18), 2783-2798.
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5

Multiparameter Flow Cytometry Analysis

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The following antibodies were used (from BD Biosciences unless otherwise noted): CD8α-Pacific Blue or AlexaFluor (AF)700 (53-6.7, Biolegend); CD4 fluoroscein isothiocyanate (FITC) (GK1.5, eBiosciences), allophycocyanin (APC) (RM4-5), phycoerythrin (PE)-Cy5 or PE-Cy7 (RM4-5, Biolegend), or PE-TexasRed (RM4-5, Invitrogen), TCRβ APC-e780 (H57-597, eBiosciences), IL4Ra PE (mIL4R-M1), CD62L APC (MEL-14),, CD44 AF700 (IM7, Biolegend), CD122-biotin (TM-1) followed by streptavidin-PE Texas Red, Eomes-PE or AF647 (Dan11mag, eBiosciences), IFNγ-PerCPCy5.5 (XMG1.2, Biolegend), APC or PE, pSTAT6 PE (J1-773.58.11), and pAkt T308 PE (J1-223.371). For pS6 staining, cells were stained with a rabbit anti-phospoS6 monoclonal (2F9; Cell Signaling) and then stained with AF488 goat anti-rabbit IgG (Invitrogen). Live/Dead Aqua (Invitrogen) was used to exclude dead cells.
Carty S.A., Koretzky G.A, & Jordan M.S. (2014). Interleukin-4 Regulates Eomesodermin in CD8+ T Cell Development and Differentiation. PLoS ONE, 9(9), e106659.
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6

Comprehensive Hematopoietic Lineage Analysis

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PB of mice was collected retro-orbitally, and hematopoietic parameters were measured by complete blood counts. Central bone marrow was flushed from femurs, and cellularity was quantified with 3% acetic acid in methylene blue (STEMCELL Technologies, 07060). For flow cytometric analysis of the lineage cell compartment of mice, flushed bone marrow cells were stained for myeloid (rat anti–Gr-1 APC-Cy7, BD Biosciences, 557661; rat anti–Mac-1 APC, eBioscience, 17-0112-83) or lymphoid lineages (rat anti-B220 FITC, eBioscience, 11-0452-82; hamster anti-CD3 PE-Cy7, eBioscience, 25-0031-82). The HSPC compartment was analyzed by staining for Lineage (biotin–Ter-119, –Mac-1, –Gr-1, -CD4, -CD8α, -CD5, -CD19, and -B220, eBioscience, 13-5921-85, 13-0051-85, 13-5931-86, 13-0112-86, 13-0452-86, 13-0041-86, 13-0081-86, 13-0193-86) followed by staining with streptavidin–PE-Texas Red (Invitrogen, SA1017), rat anti-cKit APC-Cy7 (eBioscience, 47-1171-82), rat anti-Sca1 PerCP-Cy5.5 (eBioscience, 45-5981-82), hamster anti-CD48 APC (eBioscience, 17-0481-82), and rat anti-CD150 PE-Cy7 (BioLegend, 115914). All flow cytometry analysis was performed on a BD LSR Fortessa flow cytometer with FlowJo v10.7.1 for Mac.
Zhong L., Yao L., Holdreith N., Yu W., Gui T., Miao Z., Elkaim Y., Li M., Gong Y., Pacifici M., Maity A., Busch T.M., Joeng K.S., Cengel K., Seale P., Tong W, & Qin L. (2022). Transient expansion and myofibroblast conversion of adipogenic lineage precursors mediate bone marrow repair after radiation. JCI Insight, 7(7), e150323.
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7

Multiparametric Flow Cytometry Analysis

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The following flurochrome-conjugated antibodies were used for surface staining: anti-human CD3 (Brilliant Violet 650, 1:100, OKT3, BioLegend, San Diego, CA), CD4 (PeCy7, 1:100, SK3, BioLegend), CD8 (APC-Cy7, 1:100, SK1, BD Biosciences), CD19 (PE Texas Red, 1:100, SJ25-C1, Invitrogen or APC, HIB19, Biolegend), CD25 (FITC, 1:50, 2A3, BD Biosciences, San Jose, CA), CD31 (APC, 1:100, WM59, eBioscience, San Diego, CA), CD45RA (Brilliant Violet 605, 1:100, HI100, BioLegend), CD45RO (PerCpFl710, 1:100, UCHL1, eBioscience), CD69 (Brilliant Violet 421, 1:100, FN50, BioLegend or BUV395, 1:100, FN50, BD Biosciences), CD103, (Alexa Fluor 647, 1:100, Ber-ACT8, BioLegend), CD127 (BV711, 1:100, A019D5, BioLegend), CCR7 (Alexa Fluor 488, 1:100, TG8, BioLegend). For intracellular staining, surface stained cells were resuspended and incubated in fixation buffer (eBioscience), washed, resuspended in 0.1mL permeabilization buffer (eBioscience) and stained with anti-Foxp3 antibodies (PE, 1:20, 236A/E7, eBioscience) and Ki67 (α700, 1:100, Ki-67, BioLegend) for 30 min at room temperature and washed twice with permeabilization buffer. Stained cells were acquired on a 6-laser LSRII analytical flow cytometer (BD Biosciences) in the CCTI flow cytometry core and analyzed using FlowJo software (Treestar, Ashland, OR).
Thome J.J., Bickham K.L., Ohmura Y., Kubota M., Matsuoka N., Gordon C., Granot T., Griesemer A., Lerner H., Kato T, & Farber D.L. (2015). Early life compartmentalization of human T cell differentiation and regulatory function in mucosal and lymphoid tissues. Nature medicine, 22(1), 72-77.
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