Siglecf alexa fluor 647

Manufactured by BD

SiglecF-Alexa Fluor 647 is a fluorescently-labeled antibody that binds to the SiglecF receptor. Siglec-F is a member of the sialic acid-binding immunoglobulin-like lectin (Siglec) family and is expressed on eosinophils and some macrophages. The Alexa Fluor 647 dye provides a strong fluorescent signal that can be detected using flow cytometry or other fluorescence-based techniques.

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Lab products found in correlation

Nigericin, by Merck Group (1 mentions) Ly6g bv421, by BioLegend (1 mentions) Anti mouse caspase 1 p20 ag 20b 0042, by Adipogen (1 mentions) Cd11b percp cy5, by BD (1 mentions) Cd19 fitc, by Thermo Fisher Scientific (1 mentions) Cd45 bv510, by BioLegend (1 mentions) Cd11c percp cy5, by BioLegend (1 mentions) Ly6g pe, by BioLegend (1 mentions) Cd45 pe, by Thermo Fisher Scientific (1 mentions) Cd11b pe cy7, by Thermo Fisher Scientific (1 mentions) Cd19 pe cy7, by BioLegend (1 mentions) Ultrapure lps, by Thermo Fisher Scientific (1 mentions) Anti β actin 66009 1 ig, by Proteintech (1 mentions) Anti mouse il 1β p17 af 401 na, by R&D Systems (1 mentions) Anti nlrp3 ag 20b 0014, by Adipogen (1 mentions) Cd3e pe cy7, by BD (1 mentions) Mhcii apc efluor 780, by Thermo Fisher Scientific (1 mentions) Cd11c bv510, by BioLegend (1 mentions) Cd3 fitc, by BD (1 mentions) Cd64 pe x54 5 7.1, by BioLegend (1 mentions)

3 protocols using siglecf alexa fluor 647

Inflammasome Activation Assay Protocol

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HDM was from Greer Laboratories. Anti-mouse IL-1β (p17) (AF-401-NA) was from R&D. Anti-mouse caspase-1 (p20) (AG-20B-0042) and anti-NLRP3 (AG-20B-0014) antibodies were from Adipogen. Anti-ASC (67824) antibody was from Cell Signaling Technology. Anti-β-actin (66009-1-Ig) was from Proteintech.
Anti-mouse antibodies used for flow cytometry were: CD3-FITC (BD, 553062, 145-2C11), CD19-FITC (eBioscience, 11-0193-82, 1D3), Ly-6G-PE (Biolegend, 127608, 1A8), CD45-PE (eBioscience, 12-0451-81, 30-F11), CD11c-PerCP-Cy5.5 (Biolegend, 117328, N418), CD11b-PerCP-Cy5.5 (BD, 550993, M1/70), CD11b-PE-Cy7 (eBioscience, 25-0112-82, M1/70), CD3e-PE-Cy7 (BD, 552774, 145-2C11), CD19-PE-Cy7 (Biolegend, 115520, 6D5), SiglecF-Alexa Fluor 647 (BD, 562680, E50-2440), MHCII-APC-eFluor 780 (eBioscience, 47-5321-82, M5/114.15.2), Ly-6G-BV421 (Biolegend, 127628, 1A8), CD45-BV510 (Biolegend, 103137, 30-F11), CD11c-BV510 (Biolegend, 117338, N418). Ultrapure LPS was obtained from Invitrogen. Nigericin was obtained from Sigma-Aldrich. RRx-001 (S8405) was from Selleck.

Ma M., Li G., Qi M., Jiang W, & Zhou R. (2021). Inhibition of the Inflammasome Activity of NLRP3 Attenuates HDM-Induced Allergic Asthma. Frontiers in Immunology, 12, 718779.

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Multicolor Flow Cytometry Analysis

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Commercial antibodies to the indicated murine antigens were used: CD11c-phycoerythrin (PE)/Cy7, CD11c-allophycocyanin (APC) (N418), CD11c-APCCy7 (N418), CD64-PE (X54-5/7.1), F4/80-PerCP/Cy5.5, F4/80-APC (BM8), I-A^k-PE (10-3.6), Ly-6C-PacBlue (HK1.4), Ly-6C-BV510 (HK1.4), CD45-PacBlue (30-F11), CD45-BV785 (30-F11), and Ly-6G-PacBlue (1A8) (all from BioLegend San Diego, CA); CD11b-fluorescein isothiocyanate (M1/70), CD11b-PeCy7 (M1/70), and CD11c-PE (N418) (all from eBioscience, San Diego, CA); CD45-BV510 (104), SiglecF-PE, SiglecF–Alexa Fluor 647, SiglecF-APC700, and (E50-2440) Ly6G-BUV395 (1A8) (all from BD Biosciences, San Jose, CA); and MerTK-PE/Cy7 (DS5MMER) (from Invitrogen, Carlsbad, CA). BrdU labeling was performed using the BrdU-APC labeling Kit (BD Biosciences) according to the manufacturer’s protocol. Cells were acquired either on the BD Biosciences LSRFortessa or with a BD FACScan flow cytometer with DxP multicolor upgrades by Cytek Development Inc. (Woodland Park, NJ) and then analyzed using FlowJo software (FlowJo LLC, Ashland, OR). Samples were preincubated with Fc-block [Hybridoma 2.4G2, American Type Culture Collection (ATCC)].

Todd E.M., Ramani R., Szasz T.P, & Morley S.C. (2019). Inhaled GM-CSF in neonatal mice provides durable protection against bacterial pneumonia. Science Advances, 5(8), eaax3387.

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Isolation and Flow Cytometry Analysis of Lung Leukocytes

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Lung leukocytes were isolated and labeled for analyses by flow cytometry as previously described. Briefly, mice were euthanized using a lethal dose of 1.3% tribromoethanol (Avertin, Sigma-Aldrich) and exsanguinated from the descending aorta. Lung lobes were then isolated and placed in DMEM and were cut into small pieces with a sterile razor blade and digested at 37°C for 30 min in DMEM with 2% FCS, 400 U/ml collagenase D (Roche), and 0.25 mg/ml DNAse I (Roche). Digested tissue were then homogenized by pipetting, filtered through a 100 μm filter and stained for flow cytometry. Antibodies used included Siglec-F PE (E50-2440; BD Biosciences), Siglec-F Alexa Fluor 647 (E50-2440; BD Biosciences), Ly6C Alexa Fluor^™ 647 (HK-14; eBioscience), Ly6G APC-Cy7 (1A8; Biolegend), CD45 Alexa Fluor^™ 700 (30-F11; Biolegend), CD11b PerCP-Cy5.5 (M1-70; Biolegend), CD11c PE-Cy7 (N418; eBioscience), CD4 APC-Cy7 (GK1.5; Biolegend), MerTK Biotin (Polyclonal; R&D Systems), FcεRIα APC (MAR-1; eBioscience), CD64 PE (X54-5/7.1; Biolegend), MHC-II Alexa Fluor 488 (N22), MHC-II Biotin (N22), Streptavidin eFluor 450 (eBioscience), Streptavidin APC-Cy7 (Biolegend). Flow cytometry was performed on Fortessa (BD Biosciences) and analyzed using FlowJo (Tree Star).

Sen D., Jones S.M., Oswald E.M., Pinkard H., Corbin K, & Krummel M.F. (2016). Tracking the Spatial and Functional Gradient of Monocyte-To-Macrophage Differentiation in Inflamed Lung. PLoS ONE, 11(10), e0165064.

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