Cd16 cd32 clone 2.4g2

Manufactured by BD

Sourced in United States

The CD16/CD32 (clone 2.4G2) is a laboratory reagent that binds to the Fc receptors CD16 and CD32. It is commonly used in flow cytometry and other immunological assays to block Fc-mediated non-specific binding, allowing for more accurate detection and analysis of target cells or molecules.

Automatically generated - may contain errors

Lab products found in correlation

Fixable viability dye fluortm 780, by Thermo Fisher Scientific (2 mentions) Cd90.2 clone 53 2 1, by BD (2 mentions) Diva software, by BD (2 mentions) Bovine calf serum (bcs), by Merck Group (2 mentions) Hanks balanced salt solution (hbss), by Merck Group (2 mentions) Sodium azide, by Merck Group (2 mentions) Anti cd11b clone m1 70, by Thermo Fisher Scientific (1 mentions) Anti gr 1 clone rb6 8c5, by BioLegend (1 mentions) Anti cd11c n418, by Thermo Fisher Scientific (1 mentions) Anti cd45 clone 30 f11, by Thermo Fisher Scientific (1 mentions) Moflo astrio, by Beckman Coulter (1 mentions) Fixable viability dye efluor 506, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions) Streptavidin, by BD (1 mentions) Facsaria 2 sorter, by BD (1 mentions) Cd3e clone 145 2c11, by Thermo Fisher Scientific (1 mentions) Lsrfortessa flow cytometry, by BD (1 mentions) Nk1.1 clone pk136, by BD (1 mentions) Cd117 clone 2b8, by BD (1 mentions) Ter 119 clone ter119, by BD (1 mentions)

14 protocols using cd16 cd32 clone 2.4g2

Multi-color Flow Cytometry Gating

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

Cells were acquired on a FACSCanto II (BD Bioscience, Heidelberg, Germany) and analyzed with FlowJo software (TreeStar). Cell sorting was performed on a MoFlo Astrios (Beckman Coulter, Krefeld, Germany). The following antibodies were used for staining cells: anti-CD45 (clone 30-F11, eBioscience, San Diego, USA), anti-CD11b (clone M1/70, eBioscience, San Diego, USA), anti-Gr1 (clone RB6-8C5, Biolegend, Fell, Germany), anti-Mac3 (clone M3/84 Biolegend, Fell, Germany), and anti-CD11c (N418, eBioscience San Diego, USA). Before surface staining, dead cells were stained using the Fixable Viability Dye eFluor^® 506 (eBioscience, San Diego, USA) followed by incubation with Fc receptor blocking antibody CD16/CD32 (clone 2.4G2, BD Bioscience, Heidelberg, Germany).

Hagemeyer N., Hanft K.M., Akriditou M.A., Unger N., Park E.S., Stanley E.R., Staszewski O., Dimou L, & Prinz M. (2017). Microglia contribute to normal myelinogenesis and to oligodendrocyte progenitor maintenance during adulthood. Acta neuropathologica, 134(3), 441-458.

+ Open protocol

+ Expand

Ovalbumin-specific B cell analysis

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

Draining lymph nodes (dLN; inguinal) were harvested 7 days after each immunization. Cells were stained with W614A-3S coupled with biotinylated ovalbumin (Ova) protein (Covalab) for 30 min at room temperature before cell surface antigen staining with a standard method after receptor Fc blocking with CD16/CD32 (clone 2.4G2; BD Biosciences, San Jose, CA, USA), and the following anti-mouse Abs: CD3e (clone 145-2C11; eBioscience, San Diego, CA, USA), CD45R/B220 (clone RA3-6B2), CD19 (clone 1D3), IgG1 (clone A85-1), IgD (clone 11-26c.2a), T- and B-cell activation antigen (clone GL7), and streptavidin (BD Biosciences). For cell analysis, dead cells were excluded by using the LIVE/DEAD fixable kit (Molecular Probes, Eugene, OR, USA). Cells were analyzed by BD LSRFortessa flow cytometry or isolated by BD FACSAria II sorter. A pool of five mice per condition at W3 and W5 was used for W614A-3S-specific B cell isolations (BioMark Dynamic array). A pool of 25 mice per condition at W11 was used for W614A-3S-specific IgG1⁺ GC (GL7⁺IgD^Low) and NGC (GL7^-IgD⁺) B cell isolations [chromium single cell V(D)J assay].

Bonduelle O., Chaudesaigues C., Tolazzi M., Suleiman E., de Bernard S., Alves K., Nourikyan J., Bohec M., Baudrin L.G., Katinger D., Debré P., Scarlatti G., Vieillard V, & Combadière B. (2022). Dichotomy in Neutralizing Antibody Induction to Peptide-Conjugated Vaccine in Squalene Emulsion Contrast With Aluminum Hydroxide Formulation. Frontiers in Immunology, 13, 848571.

+ Open protocol

+ Expand

Multiparametric Immune Cell Profiling

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

Two million cells were stained with Fixable Viability Dye FluorTM 780 (eBioscience, Invitrogen) to perform a live/dead staining and preïncubated with CD16/CD32 (clone 2.4G2; BD Biosciences) to block non-antigen-specific binding of Igs. Combinations of anti-mouse fluorochrome-conjugated mAbs against CD45 (clone 30-F11), CD127 (clone SB/199), CD117 (clone 2B8), CD90.2 (clone 53-2.1), KLGR-1 (clone 2F1), NK1.1 (clone PK136), CD11b (clone M1/70), CD19 (clone 1D3), CD3e (clone G4.18), CD45RB (clone 16A), CD5 (clone 53-7.3), CD94 (clone 20d5), Gr-1 (clone RB6-8C5), TCRγδ (clone GL3) and Ter-119 (clone TER-119) (all from BD Biosciences) were used to label the cells (see gating strategy in Supplementary Figs. S1 and S2). Data acquisition was performed on a LSR Fortessa flow cytometer running DIVA software (BD Biosciences). Data analysis was performed by using FlowJo software.

Pollaris L., Decaesteker T., Van den Broucke S., Jonckheere A.C., Cremer J., Verbeken E., Maes T., Devos F.C., Vande Velde G., Nemery B., Hoet P.H, & Vanoirbeek J.A. (2020). Involvement of Innate Lymphoid Cells and Dendritic Cells in a Mouse Model of Chemical-induced Asthma. Allergy, Asthma & Immunology Research, 13(2), 295-311.

+ Open protocol

+ Expand

Tumor and Immune Cell Profiling by Flow Cytometry

Cited 1 time

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

Samples were prepared for flow cytometry as described previously^{54 (link)}. Briefly, single cell suspensions were generated from tumours using the mouse Tumour Dissociation Kit (Miltenyi) and splenocytes by mechanical dissociation. Cells were stained for viability (FVS510, BD Biosciences) and incubated with an Fc-blocking antibody (CD16/CD32, clone 2.4G2, BD Biosciences) prior to extracellular staining. Extracellular staining was carried out at room temperature (RT) for 20 min, samples were fixed in 1% paraformaldehyde (PFA) and stored overnight at 4 °C until samples were acquired. For intracellular staining of IFN-γ, the Foxp3/Transcription Factor Staining Buffer Set (eBioscience™) was used following the manufacturer’s instructions. For YAC-1/NK co-culture experiments, cells were stained with Ethidium Homodimer (EthD-1, Invitrogen) to differentiate live/dead cells prior to flow cytometric analysis and were acquired on the same day. Samples were acquired on a BD LSR Fortessa or BD FACS-Celesta flow cytometer. Data were analyzed using FlowJo software (v10.8.1).

Galpin K.J., Rodriguez G.M., Maranda V., Cook D.P., Macdonald E., Murshed H., Zhao S., McCloskey C.W., Chruscinski A., Levy G.A., Ardolino M, & Vanderhyden B.C. (2024). FGL2 promotes tumour growth and attenuates infiltration of activated immune cells in melanoma and ovarian cancer models. Scientific Reports, 14, 787.

+ Open protocol

+ Expand

Profiling Mouse Myeloid Cell Populations

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

Cells were washed with phosphate-buffered saline and incubated with rat anti-mouse FCγIII/II receptor (CD16/CD32; clone 2.4G2; BD Pharmingen, San Jose, California) antibody followed by APC anti-mouse CD115/cFMS/CSF-1R antibody or matched IgG control (Biolegend, San Diego, California). Before acquisition, cells were stained with propidium iodide for gating of live cells. Analysis was performed on 100,000 events collected per sample on Gallios (Beckman Coulter, Brea, California) flow cytometer and analyzed using FlowJo (Tree Star, Ashland, Oregon).

Gabrusiewicz K., Hossain M.B., Cortes-Santiago N., Fan X., Kaminska B., Marini F.C., Fueyo J, & Gomez-Manzano C. (2015). Macrophage Ablation Reduces M2-Like Populations and Jeopardizes Tumor Growth in a MAFIA-Based Glioma Model. Neoplasia (New York, N.Y.), 17(4), 374-384.

+ Open protocol

+ Expand

Isolation and Phenotyping of Tumor-Infiltrating CD8+ and CD4+ T Cells

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

Blood samples were obtained from the tail vein of tumor-bearing mice and collected in Eppendorf tubes containing 0.5M EDTA (Fluka, Biochemika). To isolate white blood cells, red blood cells (RBCs) were lysed using ACK-Buffer (0.155M ammonium chloride, 0.01M potassium hydrogen carbonate, pH 7.2). Following a 5–7 min incubation on ice with ACK buffer, cells were resuspended in FACS buffer (PBS +2% FBS) within 96-well U-bottom plates. For optimal staining, cells were pre-incubated with FcR-block (CD16/CD32, clone 2.4G2, BD Biosciences) for 15 min at 4°C. Subsequently, cells were stained with anti-CD8α (PE-Cy7, clone 53–6.7, BD Biosciences) and anti-CD4 (PE, clone H129.19, BD Biosciences) for 20 min at 4°C. After staining, the cells underwent three washes and were resuspended in 400μL of FACS-buffer. The gating strategy on lymphocytes is illustrated in Figure S1A.

Josi R., Speiser D.E., de Brot S., Vogt A.C., Sevick-Muraca E.M., Tolstonog G.V., Bachmann M.F, & Mohsen M.O. (2024). A tetravalent nanovaccine that inhibits growth of HPV-associated head and neck carcinoma via dendritic and T cell activation. iScience, 27(4), 109439.

+ Open protocol

+ Expand

Gating Myeloid Cells via FACS

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

Peritoneal cells were resuspended in PBS containing 2 mM EDTA and 0.5% FBS. Fc receptor-mediated and nonspecific-antibody binding was blocked with CD16/CD32 (clone 2.4G2, BD Biosciences Pharmingen) for 10 min at 4°C. The cells were stained with mouse anti-GR1-FITC (1:100, BD Pharmingen) for 30 min at 4°C, and expression was analyzed by flow cytometry (FACSCalibur). Data were analyzed with WinMDi and FlowJo Version 7.6.4 software.

Sisti F., Wang S., Brandt S.L., Glosson-Byers N., Mayo L., Son Y.M., Sturgeon S., Filgueiras L., Jancar S., Wong H., Dela Cruz C.S., Andrews N., Alves-Filho J.C., Cunha F.Q, & Serezani C.H. (2018). Nuclear PTEN enhances the maturation of a microRNA regulon to limit MyD88-dependent susceptibility to sepsis. Science signaling, 11(528), eaai9085.

+ Open protocol

+ Expand

Immunophenotyping of Immune Cells in Mice

Cited 2 times

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

The spleens and thymuses of the mice from BLN-NPs, PGG-NPs, and control groups were collected, minced, and meshed on 70 μm filters. Immunofluorescence staining was performed as described in a previous study^{7 (link),8 (link)}. Briefly, cells were incubated with Fc block solution (purified anti-mouse CD16/CD32, clone 2.4G2, BD Biosciences) for 15 min at room temperature to prevent non-specific binding. For cell surface markers, cells were incubated with a fluorescently conjugated antibody against CD 68, APC anti-CD 68 (clone FA-11) (BioLegend, San Diego, CA), in the dark for 30 min at 4 °C. After washing of cells in fluorescence-activated cell sorting (FACS) buffer, flow cytometric data acquisition was performed using an Attune NxT flow cytometer (Thermo Fisher Scientific, Waltham, MA). Data analysis was performed using FlowJo (TreeStar, Ashland, OR) software.

Wang X., Parasaram V., Dhital S., Nosoudi N., Hasanain S., Lane B.A., Lessner S.M., Eberth J.F, & Vyavahare N.R. (2021). Systemic delivery of targeted nanotherapeutic reverses angiotensin II-induced abdominal aortic aneurysms in mice. Scientific Reports, 11, 8584.

+ Open protocol

+ Expand

Multicolor Flow Cytometry for Immune Profiling

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

Processed cells from each mouse were stained with Live Dead Aqua Dead Cell Kit (Invitrogen) for 30 minutes on ice, washed with PBS and then blocked with rat anti-mouse Fc antibody (CD16/CD32 clone 2.4G2, BD Biosciences) in FACs buffer for 10 minutes. Following blocking, cells were stained for the following anti-mouse fluorophores for 1 hour on ice: CD3-PerCP Cy5.5 (Biolegend), CD3-APC Cy7 (Biolegend), CD4-APC Fire (Biolegend), CD8-PE Cy7 (Biolegend), CD44-PE (Biolegend), CD62L-APC (Biolegend), CCR7-BV421 (Biolegend), CD11b-PETR (ThermoFisher), Ly6C-PerCP Cy5.5 (Biolegend), Ly6G-V450 (Biolegend), F480-PE (ThermoFisher), and CXCR4-APC (Biolegend). Cells were then washed, resuspended in FACs buffer and assayed on a Cytoflex flow cytometer (Beckman Coulter). FACs buffer consisted of HBSS (Sigma) with 2% bovine calf serum (Sigma), 0.1% sodium azide (Sigma) and 0.1% HEPES.
The “total number” of immune cells was determined by the end gated count by FACS analysis of the single cell suspension and represent the absolute number of cells infiltrating in the entire analyzed tissue. The “percentage” of immune cells represents the percentage of an immune cell subtype gated among the gated live immune cell counts as determined by Live Dead Aqua and the plot of side scatter vs forward scatter.

Blair A.B., Kim V.M., Muth S.T., Saung M.T., Lokker N., Blouw B., Armstrong T.D., Jaffee E.M., Tsujikawa T., Coussens L.M., He J., Burkhart R.A., Wolfgang C.L, & Zheng L. (2019). Dissecting the stromal signaling and regulation of myeloid cells and memory effector T cells in pancreatic cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(17), 5351-5363.

+ Open protocol

+ Expand

Multicolor Flow Cytometry for Cellular Phenotyping

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

Four million cells were stained with Fixable Viability Dye FluorTM 780 (eBioscience, Invitrogen, Merelbeke, Belgium) to perform a viability staining and pre-incubated with CD16/CD32 (clone 2.4G2, BD Biosciences, Erembodegem, Belgium) to block non-antigen-specific binding of immunoglobulins. Combinations of anti-mouse fluorochrome-conjugated mAbs against CD45 (clone 30-F11), CD127 (clone SB/199), CD90.2 (clone 53-2.1), KLRG-1 (clone 2F1), CD11b (clone M1/70), CD19 (clone 1D3), CD3e (clone G4.18), CD45RB (clone 16A), CD49b (Clone AK-7), CD5 (clone 53-7.3), TCRγδ (clone GL3), Ter-119 (clone TER-119) (all from BD Biosciences, Erembodegem, Belgium), RORγT (clone B2D)(ThermoFisher Scientific, Massachusetts, US), NKp46 (clone 29A1.4), Ly6G (Clone 1A8) and CD94 (clone 20d5)(all from Biolegend, San Diego, California) were used to label the cells. Sample acquisition was performed on a LSR Fortessa SORP flow cytometer running DIVA software (BD Biosciences, Erembodegem, Belgium). Data analysis was performed by using FlowJo software. Gating strategy available in Appendix A Fig. S4. Flow cytometry configuration are available in Appendix B.

Decaesteker T., Vanhoffelen E., Trekels K., Jonckheere A.C., Cremer J., Vanstapel A., Dilissen E., Bullens D., Dupont L.J, & Vanoirbeek J.A. (2021). Differential effects of intense exercise and pollution on the airways in a murine model. Particle and Fibre Toxicology, 18, 12.

+ 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!