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Siglecf e50 2440

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SiglecF (E50-2440) is a lab equipment product manufactured by Thermo Fisher Scientific. It is designed for use in research applications. The core function of this product is to detect and analyze the expression of the SiglecF protein in biological samples.

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Lab products found in correlation

Cd11b, by Thermo Fisher Scientific (2 mentions) Gata 3 pe, by Thermo Fisher Scientific (2 mentions) Foxp3 pe cy7, by Thermo Fisher Scientific (2 mentions) Cd3 bv605, by BioLegend (2 mentions) Cd11c pe cy7, by BioLegend (2 mentions) Cd16 32 fc block, by BD (2 mentions) Fortessa, by BD (2 mentions) Live dead fixable aqua stain, by Thermo Fisher Scientific (2 mentions) Foxp3 staining buffer kit, by Thermo Fisher Scientific (2 mentions) Ly6g bv785, by BioLegend (2 mentions) Cd11b efluor450 m1 70, by Thermo Fisher Scientific (2 mentions) Cd19 bv711 1d3, by Thermo Fisher Scientific (2 mentions) Cd4 allophycocyanin efluor780 rm4 5, by Thermo Fisher Scientific (2 mentions) Klrg1 pe efluor610 2f1, by BioLegend (2 mentions) Cd127 percp ef710 sb 199, by BioLegend (2 mentions) Cd11b allophycocyanin cy7, by BioLegend (2 mentions) Ly6c bv606, by BioLegend (2 mentions) Cd3 fitc 17a2, by Thermo Fisher Scientific (2 mentions) Cd45 bv711, by BioLegend (2 mentions) Fjk 16, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

SiglecF (17 citations) E50-2440 (3 citations)

8 protocols using siglecf e50 2440

1

Multiparametric Flow Cytometry Analysis

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Ears were collected and processed as described54 (link). All cells were first pre-incubated with anti-mouse CD16/CD32 for blockade of Fc γ receptors, then were washed and incubated for 40 min with the appropriate monoclonal antibody conjugates in a total volume of 200 μl PBS containing 2 mM EDTA and 2% (vol/vol) bovine serum. DAPI (Invitrogen) was used to distinguish live cells from dead cells during cell analysis. Stained cells were analyzed on a FACS Canto or LSRII machine using the Diva software (BD Bioscience). Data were analyzed with FlowJo software (TreeStar). The following fluorochrome-conjugated anti-mouse antibodies were used at indicated dilutions: CD103 (2E7, 1:200), CD11c (N418, 1:200), CD24 (30-F11, 1:200), CD11b (M1/70, 1:200), MHC-II (M5/114.15.2, 1:400), CD45 (30-F11, 1:200), CD64 (X54-5/7.1, 1:200), Ly6C (HK1.4, 1:200), TCRb (H57-597, 1:200), CD3 (145-2C11, 1:200), TCRgd (EBIOGL3, 1:200), B220 (RA3-6B2, 1:200), CD49b (DX5, 1:200) and FCeR1 (MAR-1, 1:200) were from eBioscience; Siglec-F (E50-2440, 1:200), Ly6G (1A8, 1:200), CD117 (2B8, 1:200), CD8 (53-6.7, 1:200), and CD4 (GK1.5, 1:200) were from BD Bioscience.
Chen L., Deshpande M., Grisotto M., Smaldini P., Garcia R., He Z., Gulko P.S., Lira S.A, & Furtado G.C. (2020). Skin expression of IL-23 drives the development of psoriasis and psoriatic arthritis in mice. Scientific Reports, 10, 8259.
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2

Flow Cytometry Analysis of Immune Cells

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To collect the non-adherent cells from the airways, BALF was harvested using phosphate-buffered saline (PBS). The lung tissues were homogenized, and the cells were then separated with Percoll gradients. Frosted glass microscope slides were used to make cell suspensions from tissues, including the MLNs and spleens. To perform the flow cytometric analysis, surface marker antibodies including CD3 (17A2), CD4 (RM4–5), CD8 (53–6.7), CD11b (M1/70), CD11c (N418), CD45 (30-F11), F4/80 (BM8), CD49b (DX4), CD69 (H1.2F3) and Siglec F (E50-2440) (eBioscience or BD Pharmingen) were used to distinguish the cell populations. To determine the antigen-specific T cell responses, cells from the BALF and lungs were (KYKNAVTEL)24 (link) and stimulated with synthetic F85–93 (WAICKRIPNKKPG)25 (link) peptides in the presence G183–195 of brefeldin A (BFA) at 37 °C for 5 hrs. Intracellular cytokine levels were measured by using monoclonal antibodies against IL-4 (BVD6-24G2), IFN-γ (XMG1.2), and TNF-α (MP6-XT22). The samples were acquired using a Becton-Dickinson LSR-II/Fortessa flow cytometer (BD, San Diego, CA) and the results were analyzed using the Flowjo software (Tree Star Inc.).
Lee Y.T., Ko E.J., Kim K.H., Hwang H.S., Lee Y., Kwon Y.M., Kim M.C., Lee Y.N., Jung Y.J, & Kang S.M. (2017). Cellular Immune Correlates Preventing Disease Against Respiratory Syncytial Virus by Vaccination with Virus-Like Nanoparticles Carrying Fusion Proteins. Journal of biomedical nanotechnology, 13(1), 84-98.
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3

Flow Cytometry Immunophenotyping

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Cells were stained with BD Biosciences (Oxford, U.K.) mAbs: CD45-PerCP-Cy5.5 (30-F11), CD45.2-V450 (104), CD4-BV650 (RM4-5), CD3-FITC (17A2), CD11b-eFluor450 (M1/70), CD19-BV711 (1D3), SiglecF (E50-2440), CD103-PE-CF594 (M290), Ly6G-BV650 (1A8); eBioscience (Loughborough, U.K.) mAbs: CD4-allophycocyanin-eFluor780 (RM4-5); Invitrogen (Dublin, Ireland) mAbs: KLRG1-PE-eFluor610 (2F1) and CD127-PerCP-ef710 (SB/199); and BioLegend (London, U.K.) mAbs: CD45-BV711 (clone: 30-F11), CD3-BV605 (17A2), CD11b-allophycocyanin-Cy7 (M1/70), CD11c-PE-Cy7 (N418), Ly6G-BV785 (1A8), Ly6C-BV606 (HK1.4), and SiglecF-allophycocyanin (S1700L). Before surface staining, Fc receptors were blocked using Fc-Block CD16/32 (BD Biosciences), and cells were incubated with LIVE/DEAD Fixable Aqua stain (Molecular Probes, Invitrogen) to isolate dead cells. For staining of transcription factors, cells were fixed and permeabilized using the Foxp3 staining buffer kit (Invitrogen) and stained with mAbs: GATA3-PE (TWAJ) and Foxp3-PE-Cy7 (FJK-16s). For the detection of YFP, along with intracellular transcription factors from Rora-YFP mice, after surface markers and viability stain, cells were prefixed with 2% paraformaldehyde followed by Foxp3 staining buffer kit. Cells were analyzed using a BD Fortessa (BD Biosciences), and data were analyzed using FlowJo software (Tree Star, Ashland, OR), using appropriate controls.
Roberts J., Chevalier A., Hawerkamp H.C., Yeow A., Matarazzo L., Schwartz C., Hams E, & Fallon P.G. (2023). Retinoic Acid–Related Orphan Receptor α Is Required for Generation of Th2 Cells in Type 2 Pulmonary Inflammation. The Journal of Immunology Author Choice, 211(4), 626-632.
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4

Comprehensive Immune Cell Profiling

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Antibodies for cell surface staining for CD4 (RM4–5), D011.10 TCR (KJ1–26), CD25 (eBio3C7), CD44 (IM7), CD62L (MEL-14), GITR (YGITR 765), Ly6G (1A8), Ly6C (HK1.4), CD45, B220, MHC Class II (M5/114.15.2), CD11b (M1/70), CD3 (2C11), Siglec-F (E50-2440) and CD11c (N418) were purchased from eBioscience, Biolegend or BD Biosciences. Unlabeled anti-CD3 (2C11) and anti-CD28 (PV-1) were purchased from the University of California San Francisco Antibody Core. Intracellular staining for Foxp3 (FJK-16s), T-bet (4B10), IFNγ (XMG1.2), IL-17A (TC11-18H10.1), IL-10 (JES5-16E3), Ki-67 (B56) (eBioscience, Biolegend or BD Biosciences) was conducted using eBioscience Foxp3 intracellular staining reagents according to manufacturer’s instructions. Intracellular staining for phospho-STAT1 (4a) (BD Biosciences), phospho-SMAD2/3 (D27F4), phospho-AKT (D9E) and phospho-S6 Ribosomal protein (D57.2.2E) (Cell Signaling Technology) was conducted as previously described48 (link). Naïve CD4+ T cells were isolated using no-touch magnetic bead purification (MACS, Miltenyi Biotec). Where indicated, cells were further sorted as described by florescent antibody labeling using FACSAria cell sorting system (BD Biosciences). Flow cytometric analysis utilized LSRII, Canto, and FACSCalibur cytometers (BD Biosciences), and data was analyzed with FlowJo Software.
Thompson L.J., Lai J.F., Valladao A.C., Thelen T.D., Urry Z.L, & Ziegler S.F. (2016). Conditioning of naïve CD4+ T cells for enhanced peripheral Foxp3 induction by non-specific bystander inflammation. Nature immunology, 17(3), 297-303.
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5

Comprehensive Immune Cell Profiling

Cited 1 time
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Antibodies for cell surface staining for CD4 (RM4–5), D011.10 TCR (KJ1–26), CD25 (eBio3C7), CD44 (IM7), CD62L (MEL-14), GITR (YGITR 765), Ly6G (1A8), Ly6C (HK1.4), CD45, B220, MHC Class II (M5/114.15.2), CD11b (M1/70), CD3 (2C11), Siglec-F (E50-2440) and CD11c (N418) were purchased from eBioscience, Biolegend or BD Biosciences. Unlabeled anti-CD3 (2C11) and anti-CD28 (PV-1) were purchased from the University of California San Francisco Antibody Core. Intracellular staining for Foxp3 (FJK-16s), T-bet (4B10), IFNγ (XMG1.2), IL-17A (TC11-18H10.1), IL-10 (JES5-16E3), Ki-67 (B56) (eBioscience, Biolegend or BD Biosciences) was conducted using eBioscience Foxp3 intracellular staining reagents according to manufacturer’s instructions. Intracellular staining for phospho-STAT1 (4a) (BD Biosciences), phospho-SMAD2/3 (D27F4), phospho-AKT (D9E) and phospho-S6 Ribosomal protein (D57.2.2E) (Cell Signaling Technology) was conducted as previously described48 (link). Naïve CD4+ T cells were isolated using no-touch magnetic bead purification (MACS, Miltenyi Biotec). Where indicated, cells were further sorted as described by florescent antibody labeling using FACSAria cell sorting system (BD Biosciences). Flow cytometric analysis utilized LSRII, Canto, and FACSCalibur cytometers (BD Biosciences), and data was analyzed with FlowJo Software.
Thompson L.J., Lai J.F., Valladao A.C., Thelen T.D., Urry Z.L, & Ziegler S.F. (2016). Conditioning of naïve CD4+ T cells for enhanced peripheral Foxp3 induction by non-specific bystander inflammation. Nature immunology, 17(3), 297-303.
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6

Flow Cytometry Immunophenotyping

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Cells were stained with BD Biosciences (Oxford, U.K.) mAbs: CD45-PerCP-Cy5.5 (30-F11), CD45.2-V450 (104), CD4-BV650 (RM4-5), CD3-FITC (17A2), CD11b-eFluor450 (M1/70), CD19-BV711 (1D3), SiglecF (E50-2440), CD103-PE-CF594 (M290), Ly6G-BV650 (1A8); eBioscience (Loughborough, U.K.) mAbs: CD4-allophycocyanin-eFluor780 (RM4-5); Invitrogen (Dublin, Ireland) mAbs: KLRG1-PE-eFluor610 (2F1) and CD127-PerCP-ef710 (SB/199); and BioLegend (London, U.K.) mAbs: CD45-BV711 (clone: 30-F11), CD3-BV605 (17A2), CD11b-allophycocyanin-Cy7 (M1/70), CD11c-PE-Cy7 (N418), Ly6G-BV785 (1A8), Ly6C-BV606 (HK1.4), and SiglecF-allophycocyanin (S1700L). Before surface staining, Fc receptors were blocked using Fc-Block CD16/32 (BD Biosciences), and cells were incubated with LIVE/DEAD Fixable Aqua stain (Molecular Probes, Invitrogen) to isolate dead cells. For staining of transcription factors, cells were fixed and permeabilized using the Foxp3 staining buffer kit (Invitrogen) and stained with mAbs: GATA3-PE (TWAJ) and Foxp3-PE-Cy7 (FJK-16s). For the detection of YFP, along with intracellular transcription factors from Rora-YFP mice, after surface markers and viability stain, cells were prefixed with 2% paraformaldehyde followed by Foxp3 staining buffer kit. Cells were analyzed using a BD Fortessa (BD Biosciences), and data were analyzed using FlowJo software (Tree Star, Ashland, OR), using appropriate controls.
Roberts J., Chevalier A., Hawerkamp H.C., Yeow A., Matarazzo L., Schwartz C., Hams E, & Fallon P.G. (2023). Retinoic Acid–Related Orphan Receptor α Is Required for Generation of Th2 Cells in Type 2 Pulmonary Inflammation. The Journal of Immunology Author Choice, 211(4), 626-632.
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7

Multi-Marker Immunophenotyping of Immune Cells

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Single-cell suspensions of organs were treated with Gey's solution where appropriate, and washed with PBS/3% FCS. The cells were preincubated with 20 µg anti-CD16/32 antibody (clone 2.4G2), washed and stained with a combination of fluorochrome-labelled monoclonal antibodies (all steps at 4°C). Antibodies were from BD Biosciences {name [alternative name] clone)} {TCRβ (H57-597); CD4 (RM4-5 or GK1.5), CD5 ([Ly-1] 53–7.3), CD8 (53–6.7), CD11b (M1/70); CD11c (HL3), CD11c (IM7), CD19 (1D3), Siglec F (E50-2440), NK1.1 (PK136), Ly6G (1A8)}, eBioscience {I-Ab [MHC class II] AF6-120.1), H-2Kb [MHC class I] AF6-88.5.5.3), CD3e (eBio500A2 or 145-2C11), CD25 (PC61.5), CD27 (LG.7F9), CD45 (30-F11), CD90.2 (53–2.1), F4/80 (BM8), Ly6C (HK1.4)}, and MD Bioproducts {T1-ST2 (DJ8)}. Samples were processed on a LSR II flow cytometer equipped for 10-parameter acquisition, and analysed with TreeStar FlowJo software (version 9.5).
Scheer S., Krempl C., Kallfass C., Frey S., Jakob T., Mouahid G., Moné H., Schmitt-Gräff A., Staeheli P, & Lamers M.C. (2014). S. mansoni Bolsters Anti-Viral Immunity in the Murine Respiratory Tract. PLoS ONE, 9(11), e112469.
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8

Multiparameter Flow Cytometry Analysis of Lung Cells

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BALF was collected by three tracheal instillations and aspirations of 1 mL PBS. Tissues were digested with collagenase D (1.5 U/mL) and dispase II (2.4 U/mL) (Roche) for 30 minutes. Samples were then incubated with anti-CD16/32 to block nonspecific antibody binding. Fluorescence-conjugated antibodies to CD45 (I3/2), CD11c (N418), CD3e (145-2C11), CD8 (53.67), CD4 (GK1.5), B220 (RA3-6B2), Ly6B (7/4) (Abcam), SiglecF (E50-2440) (eBiosciences), CD34 (RAM34) (eBiosciences), CD31 (390) (eBiosciences), PDGFRα (APA5) (eBiosciences), Sca1 (D7) (eBiosciences), and EpCAM (G8.8) (eBiosciences) were used. For EdU uptake experiments, mice were given 1 mg EdU daily by intraperiotenal injections; EdU detection was performed using the Click-IT assay kit (Life). Data was acquired on a BD LSRII and analyzed with FlowJo Software.
All antibodies were generated in-house (UBC AbLab) unless otherwise indicated.
Lo B.C., Gold M.J., Scheer S., Hughes M.R., Cait J., Debruin E., Chu F.S.F., Walker D.C., Soliman H., Rossi F.M., Blanchet M.R., Perona-Wright G., Zaph C, & McNagny K.M. (2017). Loss of Vascular CD34 Results in Increased Sensitivity to Lung Injury. American journal of respiratory cell and molecular biology, 57(6).
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