Siglecf

Manufactured by Thermo Fisher Scientific

Sourced in United States

SiglecF is a cell surface receptor expressed on eosinophils. It functions in the regulation of eosinophil survival and trafficking.

Automatically generated - may contain errors

Lab products found in correlation

17 protocols using siglecf

Comprehensive Mouse Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies purchased from BioLegend: NK1.1 (PK136), CD11c (N418), CD19 (6D5), CD25 (PC61), TCRγ/δ (GL3), CD80 (16–10A1), CD45.1 (A20), CD45.2 (104), CD45R (B220; RA3–6B2), CD90.1 (OX-7), CD172a (SIRPα; P84), CD301b (MGL2; URA-1), CD317 (PDCA-1, BST2; 129C1), TCRβ (H57–597), XCR1 (ZET), CD11b (M1/70), Ep-CAM (G8.8), Ly-51 (6C3), Ly-6D (49-H4), CD64 (X54–5/7.1), Ly-6G (1A8), TER-119 (TER-119), CD3 (17A2).
Antibodies purchased from BD Biosciences: CD4 (GK1.5), CD8a (53–6.7), CD69 (H1.2F3), H-2K^b (AF6–88.5), TCRβ (H57–597), CD86 (GL1), CD90.2 (30-H12), Siglec-F (E50–2440), CD25 (PC61.5), CD11c (HL3)
Antibodies purchased from Thermo Fischer: CD5 (53–7.3), MHC Class II– I-A/I-E (M5/114.15.2), CCR7 (4B12), Ly-6C (HK1.4), FOXP3 (FJK-16s).
Antibodies purchased from Santa Cruz Biotechnology: MHC Class II I-Ab bound to invariant chain (li) (15G4).
Antibodies purchased from Vector Laboratories: UEA1 (Catalog # FL-1061)
H2-DO (Mags.Ob3) and H2-DM (2C3A) antibodies were a generous gift from Dr. Lisa Denzin.
Additional details are provided in Supplementary Table.

Breed E.R., Vobořil M., Ashby K.M., Martinez R.J., Qian L., Wang H., Salgado O.C., O’Connor C.H, & Hogquist K.A. (2022). Type 2 cytokines in the thymus activate Sirpα+ dendritic cells to promote clonal deletion. Nature immunology, 23(7), 1042-1051.

+ Open protocol

+ Expand

Liver Immune Cell Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse liver mononuclear cells (MNCs) and Kupffer cells were labeled with fluorescence-tagged antibodies; eFluor 450-CD45 (clone 30-F11), FITC-F4/80 (clone BM8), APC-conjugated Ly-6G (clone 1A8), PE-CF594-conjugated SiglecF (clone E50-2440), APC-Cy7-conjugated CD11b (clone M1/70), PE-conjugated CD146 (clone ME-9F1). Fortessa™ cell analyzer and FlowJo software v10.8.1 (BD bioscience, San Jose, CA, USA) were used to identify each cell based on following markers: macrophages (F4/80⁺CD11b⁺), Kupffer cells (F4/80^hiCD11b⁺), Eosinophils (CD11b⁺SiglecF⁺), neutrophils (CD11b⁺Ly6G⁺) in CD45+ cells, Live cells were determined with LIVE/DEAD™ fixable aqua dead cell stain kit (Thermo Fisher Scientific).

Kim K., Kim M.H., Kang J.I., Baek J.I., Jeon B.M., Kim H.M., Kim S.C, & Jeong W.I. (2023). Ginsenoside F2 Restrains Hepatic Steatosis and Inflammation by Altering the Binding Affinity of Liver X Receptor Coregulators. Journal of Ginseng Research, 48(1), 89-97.

+ Open protocol

+ Expand

Analyzing Immune Cell Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols

At experimental endpoints, mice were sacrificed, and BALF was collected. Cells were isolated for flow cytometry and extracellularly stained as either macrophages or neutrophils and for their activation. Antibodies (Siglec-F, Ly6G, F4/80 and CD11b) and Pacific Blue for viability were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Shipman J.G., Onyenwoke R.U, & Sivaraman V. (2024). Vaping-Dependent Pulmonary Inflammation Is Ca2+ Mediated and Potentially Sex Specific. International Journal of Molecular Sciences, 25(3), 1785.

+ Open protocol

+ Expand

Gene expression analysis of murine lung

Check if the same lab product or an alternative is used in the 5 most similar protocols

The Qiagen RNeasy Mini Kit was used to isolate RNA according to the manufacturer's instructions, after tissues were initially homogenized in TRIzol (BD Biosciences, Hercules, CA). Purified RNA was reverse transcribed by iScript RT (Bio‐Rad) into cDNA. Differences in relative gene expression were quantified using FAM‐conjugated TaqMan Gene Expression Assay (Thermo Fisher, Middletown, VA, USA). The following genes were examined by RT‐PCR: Siglecf (Mm00523987_m1; Thermo Fisher), Prg2 (Mm0043905_m1), Ccr3 (Mm00515543_s1), Emr1 (Mm00802529_m1), Tnfa (Mm00443258_m1), Itgax (Mm00498698_m1), Arg1 Mm01190441_g1), Mmp9 (Mm00442991_m1), Vegfa (Mm01281449_m1), Fgf2 (Mm00433287_m1), and Gapdh (Mm99999915_g1). Data were normalized to the housekeeping gene, Gapdh, and analyzed by the Pfaffl method (Pfaffl 2001).

Bolus W.R., Kennedy A.J, & Hasty A.H. (2018). Obesity‐induced reduction of adipose eosinophils is reversed with low‐calorie dietary intervention. Physiological Reports, 6(22), e13919.

+ Open protocol

+ Expand

Quantitative Analysis of Siglec Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Quantitative reverse-transcription PCR was used to determine relative mRNA levels in macrophages. Where stated, BMDM were stimulated with LPS at 1 μg/mL overnight. RNA extraction was done using a GeneJET RNA Purification Kit (ThermoFisher Scientific). cDNA was made using SuperScript™ VILO™ (ThermoFisher Scientific) reverse transcriptase. Taq-man probes used for CD22, CD33, Siglec-E, Siglec-F, Siglec-G, and Abt1 were from Thermo Scientific.

Imbert P.R., Saric A., Pedram K., Bertozzi C.R., Grinstein S, & Freeman S.A. (2020). An acquired and endogenous glycocalyx forms a bidirectional ‘don’t eat’ and ‘don’t eat me’ barrier to phagocytosis. Current biology : CB, 31(1), 77-89.e5.

+ Open protocol

+ Expand

Immune Cell Isolation from Murine Lungs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bronchoalveolar lavage fluids (BALF) were harvested using PBS to collect non-adherent cells from the lung airways. After collecting BALF, lung tissues were homogenated and cells were then passed through strainer and spun on 44/67% Percoll gradients at 2800 rpm for 20 minutes. A lung cell band which contained most immune cells was harvested. The frosted glass microscope slides were used to make cell suspensions from the mediastinal lymph nodes (MLN) and spleens. Red blood cells were lysed with ammonium chloride and lymphocytes were filtered through cell strainers. Flow cytometric analysis were carried out using cell surface marker antibodies specific for CD3, CD4, CD8, CD11b, CD11c, CD45, F4/80, Siglec F and Ly6c (eBioscience or BD Pharmingen). The Becton-Dickinson LSR-II/Fortessa flow cytometer (BD, San Diego, CA) was used to analyze distinct populations from the tissues and acquired samples were further analyzed by using Flowjo software (Tree Star Inc.).

Lee Y.T., Kim K.H., Hwang H.S., Lee Y., Kwon Y.M., Ko E.J., Jung Y.J., Lee Y.N., Kim M.C, & Kang S.M. (2015). Innate and adaptive cellular phenotypes contributing to pulmonary disease in mice after respiratory syncytial virus immunization and infection. Virology, 485, 36-46.

+ Open protocol

+ Expand

Multiparameter Flow Cytometry of Lung Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Bronchoalveolar lavage was performed, and cells were removed via centrifugation. Cells were stained then fixed in IC fixation buffer (eBioscience, 00-8222-49, San Diego, CA) and run and identified via Zombie NIR (Biolegend, 423105, San Diego, CA), CD45, Alexa Fluor 532 (eBioscience, 58-0459-42, San Diego, CA), CD11c, PE-Cy7 (Biolegend, 117317, San Diego, CA), CD11b, BV480 (BD Biosciences, 566117, San Jose, CA), SIGLEC F, AF700 (eBioscience, 56-1702-80, San Diego, CA), Ly-6c, FITC (Biolegend, 128005, San Diego, CA), and Ly-6G, BV650 (Biolegend, 127641, San Diego, CA) on a Cytek Aurora Borealis at the University of Virginia flow cytometry core. Neutrophils are Zombie NIR⁻, CD45⁺, CD11c⁻, CD11b⁺, and Ly-6G⁺; eosinophils are Zombie NIR⁻, CD45⁺, CD11c⁻, CD11b⁺, and SIGLEC F⁺; inflammatory monocytes are Zombie NIR⁻, CD45⁺, CD11c⁻, CD11b⁺, Ly-6G⁻, and Ly-6C⁺⁺. Data analysis and figure generation were performed in Omiq and Graphpad Prism. A two-tailed Student’s t test was used to determined statistical significance.

Moreau G.B., Burgess S.L., Sturek J.M., Donlan A.N., Petri WA J.r, & Mann B.J. (2020). Evaluation of K18-hACE2 Mice as a Model of SARS-CoV-2 Infection. The American Journal of Tropical Medicine and Hygiene, 103(3), 1215-1219.

+ Open protocol

+ Expand

Multiparametric Phenotyping of Lung Immune Cells

Cited 3 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

BALFs were harvested from the lung airways by infusion with PBS via trachea using a 25-gauge catheter (26 (link)). Lung tissues were homogenized, passed through a cell strainer, and spun on 44/67% Percoll gradients to collect lung cells. Cellular phenotypes were determined by flow cytometric analysis using cell surface marker Abs specific for CD3, CD4, CD8, CD11b, CD11c, CD45, F4/80, Siglec F, DX5, and Ly6c (eBioscience, San Diego, CA, USA or BD Pharmingen, San Jose, CA, USA) as previously described (17 (link)27 (link)28 (link)). Cells from BALF and lungs were stimulated with the synthetic F_85-93 (KYKNAVTEL) peptide (29 (link)) for CD8 T cell activation. Intracellular cytokine-producing cells stained with monoclonal IFN-γ and TNF-α and cell phenotypic marker Abs were acquired by the Becton-Dickinson LSR-II/Fortessa flow cytometer and data analyzed by Flowjo software (Tree Star Inc., Ashland, OR, USA). In the determination of cellular phenotypes, the fraction (%) of each cell phenotypes obtained by flow cytometry was multiplied by the total cell numbers counted in the cell preparations from the lung tissues and BALF.

Kwon Y.M., Hwang H.S., Lee Y.T., Kim K.H., Lee Y., Kim M.C., Lee Y.N., Quan F.S., Moore M.L, & Kang S.M. (2019). Respiratory Syncytial Virus Fusion Protein-encoding DNA Vaccine Is Less Effective in Conferring Protection against Inflammatory Disease than a Virus-like Particle Platform. Immune Network, 19(3), e18.

+ Open protocol

+ Expand

Bronchoalveolar Lavage Cell Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

For bronchoalveolar lavage (BAL) cells, the diaphragms were dissected to allow free lung expansion and the lungs were lavaged by slowly instilling 1 ml of PBS and then gently aspirating as described ^{22 (link)}. To perform cell phenotype analysis, immune cell surface marker antibodies including CD3, CD11b, CD11c, CD45, F4/80, MHC class II, and Siglec F (eBioscience or BD Pharmingen) were used to distinguish cell populations.

Lee Y.N., Hwang H.S., Kim M.C., Lee Y.T., Kim Y.J., Lee F.E, & Kang S.M. (2015). Protection against Respiratory Syncytial Virus by Inactivated Influenza Virus Carrying a Fusion Protein Neutralizing Epitope in a chimeric hemagglutinin. Nanomedicine : nanotechnology, biology, and medicine, 12(3), 759-770.

+ Open protocol

+ Expand

Lung Cell Isolation and Flow Cytometric Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lung cell isolation and flow cytometric analysis were performed as described previously (45 (link)). Briefly, lung single-cell suspensions were obtained from minced lung tissue and subjected to digestion with collagenase/dispase (1 mg/ml; Roche) and DNase I (100 µg/ml; grade II; Roche), then treated with RBC lysis buffer to remove red blood cells. Cells were suspended in PBS with 1% BSA, blocked with 2.4G2 (anti-CD16/CD32, Multi Sciences, Hangzhou, China), and labeled with specific antibodies. Isolation of lung lymphocytes for detection of NK cells was performed as described (46 (link)). Antibodies used included: SiglecF, Ly6C, CD19, CD4, CD8 (eBioscience, San Diego, CA, USA), CD45, Ly6G, CD11c, CD11b, CD3e, NK1.1 (Multi Sciences), CD107a, IFN-γ (BD Biosciences). Flow cytometry was used to analyze specific cell populations in mouse lungs using the following gates: AMs as SiglecF⁺CD11c⁺, eosinophils as SiglecF⁺CD11c^-, tissue monocytes as SiglecF^- Ly6G^-CD11b⁺Ly6C⁺, and neutrophils as Ly6G⁺CD11b⁺.

Chen B., Chen Y., Rai K.R., Wang X., Liu S., Li Y., Xiao M., Ma Y., Wang G., Guo G., Huang S, & Chen J.L. (2021). Deficiency of eIF4B Increases Mouse Mortality and Impairs Antiviral Immunity. Frontiers in Immunology, 12, 723885.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!