Anti cx3cr1 fitc

Manufactured by BioLegend

Anti-CX3CR1-FITC is a fluorescently-labeled antibody that targets the CX3CR1 receptor. CX3CR1 is a chemokine receptor involved in cell migration and adhesion processes. This product can be used for the detection and analysis of CX3CR1-expressing cells in flow cytometry applications.

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CX3CR1 (26 citations) CX3CR1-FITC (6 citations) Anti-CX3CR1 (6 citations)

5 protocols using anti cx3cr1 fitc

Characterization of DRG Myeloid Cells

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DRG single-cell suspensions were blocked for non-specific antibody-binding with anti-CD16/32 Fc-Block (BioLegend 101320, 1:100) and incubated in an antibody cocktail consisting of anti-CD163-PE (BioLegend 156703, 1:200), anti-TLR4-PE/Cy7 (BioLegend 117609, 1:200), anti-CD206-PerCP/Cy5.5 (BioLegend 141715, 1:200), anti-CD11b-APCCy7 (BioLegend 101225, 1:200), anti-CD68-Alexa700 (BioLegend 137025, 1:200), anti-CD80-Bv421 (BioLegend 104725, 1:200), anti-CD86-Bv510 (BioLegend 105039, 1:200), anti-Gr1-Bv605 (BioLegend 10844, 1:200), anti-F4/80-Bv785 (BioLegend 123141, 1:200), and anti-CX3CR1-FITC (BioLegend 149019, 1:200) for 20 min at 4 °C. Exclusion of dead cells was achieved by resuspending cells in FACS buffer containing 10 nM TO-PRO-3 (Invitrogen, Carlsbad, CA, USA, T3605). Flow cytometry from the samples was performed using a BD LSRFortessa device and BD FACSDiva Software (BD Biosciences). Raw data were analyzed using the CytoExplorer R package version 1.1.0 [46 ].

Choconta J.L., Labi V., Dumbraveanu C., Kalpachidou T., Kummer K.K, & Kress M. (2023). Age-related neuroimmune signatures in dorsal root ganglia of a Fabry disease mouse model. Immunity & Ageing : I & A, 20, 22.

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Multiparametric Flow Cytometry Analysis

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The following mouse anti-human monoclonal antibodies (mAb) were used for flow cytometry: anti-CD3-Alexa Fluor (AF) 647, (Biolegend, San Diego, CA, clone UCHT1), anti-CD8-peridinin chlorophyll(PerCP), (Biolegend, San Diego, CA, clone SK1), anti-CD8-phycoerytrin-(PE)-cyanine(Cy)7,(eBioscience Inc, San Diego, CA, clone rpa-t8), anti-TCF-1-AF 647, (Cell Signaling, Boston, MA, clone 7FIA10), anti-CX3CR1-FITC, (Biolegend, San Diego, CA, clone 2A9-1), anti-PD-1-FITC, (Becton Dickinson Bioscience, San Jose, CA, clone MIH4), anti-CD127-AF 647, (Biolegend, San Diego, CA, clone, A019D5), anti-Glut1-AF488, (R&D Systems, Minneapolis, MN, clone 202915), anti-PGC1α-AF488, (Santa Cruz Biotechnology, Santa Cruz, CA, Clone D-5), anti-CPT1a-AF488 (Abcam, UK, clone 8F6AE9), anti-P-rpS6-AF488 (Cell Signaling, Boston, MA, clone D57.2.2E), anti-CD107a-AF488, (Biolegend, San Diego, CA, clone H4A3), anti-IFNγ FITC, (R&D Systems, Minneapolis, MN, clone RMMA-1), anti-TNFα AF647 (Biolegend, San Diego, CA, clone MAb11).

Peña-Asensio J., Calvo-Sánchez H., Miquel J., Sanz-de-Villalobos E., González-Praetorius A., Torralba M, & Larrubia J.R. (2023). IL-15 boosts activated HBV core-specific CD8+ progenitor cells via metabolic rebalancing in persistent HBV infection. iScience, 27(1), 108666.

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Multiparameter Flow Cytometry Analysis

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Cells from lung, spleen and lymph nodes were incubated with Fc blocker for 10 min at 4 °C, and then stained with viability dye eFluor 780 (eBioscience), anti-CD45-PerCP-Cy5.5, anti-CD11b-APC, anti-F4/80-PE, anti-Ly6C-PerCP-Cy5.5, anti-Ly6G-PE, anti-CX3CR1-FITC, anti-CD4-APC, and anti-CD8-FITC (BioLegend; 1:300 vol/vol dilution) for 30 min at 4 °C. For BM phenotyping, cells were stained for anti-CD19-APC, anti-Ter119-APC, anti-CD11b-APC, anti-Ly6C/G-APC and anti-CD3-APC as lineage markers along with anti-Ly6A/E-APC-Cy7 (Sca-1), anti-CD117-PE-Cy7 (c-kit), anti-CD48-FITC and anti-CD150-PE-Cy5 (SLAM; BioLegend; 1:300 dilution) for Lin⁻Sca-1⁺c-kit⁺ LSK populations and Lin⁻Sca-1⁺c-kit⁺ CD48⁺CD150⁻ MPPs. Data were collected using FACSCanto flow cytometer (BD Bioscience). The intracellular TNF (anti-TNF-PE; 1:600 dilution) and FoxP3 (anti-FoxP3-PE; 1:300 dilution) staining was performed using Fixation Buffer (BioLegend, 420801) and Foxp3 Staining Buffer Set (Thermo Fisher, 00-5523-00) according to the manufacturer’s instruction. Information for all flow antibodies is provided in the Reporting Summary. Flow data were analyzed using FlowJo software (Tree Star).

Ding C., Shrestha R., Zhu X., Geller A.E., Wu S., Woeste M.R., Li W., Wang H., Yuan F., Xu R., Chariker J.H., Hu X., Li H., Tieri D., Zhang H.G., Rouchka E.C., Mitchell R., Siskind L.J., Zhang X., Xu X.G., McMasters K.M., Yu Y, & Yan J. (2023). Inducing trained immunity in pro-metastatic macrophages to control tumor metastasis. Nature immunology, 24(2), 239-254.

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Phenotypic Characterization of Immune Cells

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Antibodies used for cell labeling of isolated human monocytes, NK cells, T cells, in vitro generated TAM, M1-like, or M2-like macrophages, or single cell suspensions isolated from tumor or normal human tissue samples were as follows: V450 anti-CD14 (BD Biosciences, 560,349), PE anti-CD56 (Beckman Coulter, IM2073U), PE anti-CD3 (Beckman Coulter, IM1282), PE anti-CD19 (BD Biosciences, 555,413), PE-CF594 anti-CD163 (BD Biosciences, 562,670), PE/Cy7 anti-CD45 (BioLegend, 304,016), FITC anti-CX3CR1 (BioLegend, 341,605), PE anti-CD163 (BioLegend, 333,605), APC anti-CD206 (BD Biosciences, 561,783), and PerCP/Cy5.5 anti-PD-L1 (BioLegend, 329,737). Flow cytometry was performed on a LSR II flow cytometer.

Benner B., Scarberry L., Suarez-Kelly L.P., Duggan M.C., Campbell A.R., Smith E., Lapurga G., Jiang K., Butchar J.P., Tridandapani S., Howard J.H., Baiocchi R.A., Mace T.A, & Carson WE I.I.I. (2019). Generation of monocyte-derived tumor-associated macrophages using tumor-conditioned media provides a novel method to study tumor-associated macrophages in vitro. Journal for Immunotherapy of Cancer, 7, 140.

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Brain Tissue Cell Isolation and Characterization

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Cells were isolated from brain tissue as previously described [20 (link)], with some modifications. Briefly, isolated tissues were cut up into fine pieces and were dissociated in buffer containing HBSS (without calcium/magnesium), 5% FBS, 10 μM HEPES, 2 mg/mL collagenase D (Sigma-Aldrich) and 28U/mL DNaseI (NEB) at 37 °C for 45 min. Every 15 min, dissociated tissue was pipetted up and down using sequentially smaller Pasteur pipettes to homogenize tissue debris. Homogenate was filtered through a 70-μm filter and centrifuged 10 min at 300 g. Pellets were resuspended in PBS and stained with an APC-Cy7 LIVE/DEAD™ fixable dead cell stain kit (Invitrogen). Cells were washed, resuspended in FACS buffer with Fc block (BD Biosciences) and stained with the following antibodies: AF700 anti-CD3 (BioLegend, clone:17A2), BV711 anti-Ly6C (BioLegend, clone: 1A8), BV421 anti-Ly6G (BioLegend, clone: HK1.4), FITC anti-CX3CR1 (BioLegend, clone: SA011F11), BV510 anti-CD19 (BioLegend, clone: 6D5), PerCPCy5.5 anti-CD45 (BioLegend, clone: 30-F11), and APC anti-Cd11b (BioLegend, clone: M1/70). Samples were measured on an LSRFortessa (BD Biosciences) and analyzed using FlowJo software.

Talley S., Valiauga R., Anderson L., Cannon A.R., Choudhry M.A, & Campbell E.M. (2021). DSS-induced inflammation in the colon drives a proinflammatory signature in the brain that is ameliorated by prophylactic treatment with the S100A9 inhibitor paquinimod. Journal of Neuroinflammation, 18, 263.

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