Fcblock clone 93

Manufactured by BioLegend

Sourced in United States

FcBlock (clone 93) is a recombinant monoclonal antibody product designed to block Fc receptor interactions. It is intended for use in flow cytometry, ELISA, and other immunoassay applications.

Automatically generated - may contain errors

Lab products found in correlation

Lsrii flow cytometer, by BD (2 mentions) Facsaria 2, by BD (1 mentions) Fixable viability stain 510, by BD (1 mentions) Anti cd44 pe cy7 clone im7, by BioLegend (1 mentions) Anti cd8α apc clone 53 6, by BD (1 mentions) Zombie aqua fixable viability dye, by BioLegend (1 mentions) Facscanto 2, by BD (1 mentions) Ack lysing buffer, by Thermo Fisher Scientific (1 mentions)

4 protocols using fcblock clone 93

Multiparameter Flow Cytometry Analysis of Mouse Splenocytes

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Mouse splenocytes were prepared as described above. Samples were resuspended in PBS with 1% fetal calf serum, followed by incubation with anti-CD16/32 mAb (Fc block, clone 93, BioLegend) for 10 min at 4 °C to prevent nonspecific Fc receptor binding. Samples were then stained for 40 min at 4 °C with various combinations of fluorochrome-conjugated antibodies: anti-CD45-PerCP (clone 30-F11, BioLegend), anti-CD8α-APC (clone 53–6.7, BD Biosciences), anti-CD3-Alexa Fluor 488 (clone 145-2C11, BD Biosciences), anti-CD44-PE-Cy7 (clone IM7, BioLegend), and anti-CD62L-BV421 (clone MEL-14, BioLegend). To exclude dead cells from the analysis, a fixable viability stain 510 (BD Biosciences) was used. Samples were analyzed using BD FACS Aria II and the collected data was analyzed using FlowJo software (Tree Star).

Okada H., Takahashi K., Yaku H., Kobiyama K., Iwaisako K., Zhao X., Shiokawa M., Uza N., Kodama Y., Ishii K.J, & Seno H. (2022). In situ vaccination using unique TLR9 ligand K3-SPG induces long-lasting systemic immune response and synergizes with systemic and local immunotherapy. Scientific Reports, 12, 2132.

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Flow Cytometry Immunophenotyping of Hematopoietic Cells

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ER-Hoxb8 immortalized progenitors and SCF-dependent neutrophil progenitors were kept on ice for all steps if not stated otherwise. Cells were washed and resuspended in PBS and stained with Zombie Aqua fixable viability dye (BioLegend 423101) to exclude dead cells. Cells were washed with PBS and incubated with FcBlock (clone 93, BioLegend) for 15 min at 4 °C, followed by staining with fluorescently conjugated antibodies for 25 min at 4 °C in staining buffer (0.5% BSA in PBS). The cells were washed with staining buffer and analyzed on a LSRII flow cytometer (BD). Following cell populations were identified based on cell marker expression: ER-Hoxb8 immortalized progenitors (CD45+ CD11b− cKit/CD117+), neutrophils (CD45+ CD11b+ cKit/CD117− Ly-6G+). Antibodies are reported in Supplementary Table 2.

Ko J., Wilkovitsch M., Oh J., Kohler R., Bolli E., Pittet M.J., Vinegoni C., Sykes D.B., Mikula H., Weissleder R, & Carlson J.C. (2022). Spatiotemporal multiplexed immunofluorescence imaging of living cells and tissues with bioorthogonal cycling of fluorescent probes. Nature biotechnology, 40(11), 1654-1662.

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Murine Spleen Cell Isolation and Immunophenotyping

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Mice spleens were collected and macerated in 3.5 mL ice-cold DMEM with 10% v/v FBS and centrifuged at 1200 rpm, 4 °C, for 10 min. Erythrocytes were lysed using ACK Lysing Buffer (Gibco) according to the manufacturer protocols. Cells were counted under optical microscopy and 2 × 10⁶ cells per well were seeded in a 96-well plate in 200 μL per well. Single-cell suspensions from the spleen were incubated with 2.5 g/mL Fc block (clone 93, Biolegend, San Diego, CA, USA) to block non-specific binding for 10 min at room temperature.
Cells were stained with monoclonal antibodies anti-CD3 APC (17A2), CD4 PE (GK1.5), CD8 FITC (53-6.7), CD11b PE-Cy7 (M1/70), CD11c APC-Cy7 (N418), CD25 APC-Cy7 (PC61), CD62L FITC (MEL-14), CD69 PE-Cy7 (H1.2F3), F4/80 PE (BM8), Ly6G APC (1A8), IA/IE Percp-Cy5.5 (M5/114.15.2), all from BioLegend (San Diego, CA, USA), for 30 min in FACS Buffer at 4 °C, washed and resuspended in paraformaldehyde 2% before acquisition on the flow cytometer (FACS Canto II; BD Biosciences, San Diego, CA, USA). Data were analyzed using FlowJo software (BD Bioscience).

de Carvalho A.C., Dias C.S., Coimbra L.D., Rocha R.P., Borin A., Fontoura M.A., Carvalho M., Proost P., Nogueira M.L., Consonni S.R., Sesti-Costa R, & Marques R.E. (2023). Characterization of Systemic Disease Development and Paw Inflammation in a Susceptible Mouse Model of Mayaro Virus Infection and Validation Using X-ray Synchrotron Microtomography. International Journal of Molecular Sciences, 24(5), 4799.

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Profiling Lung Tumor Neutrophil Subsets

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Single cell suspensions from lung tumor or tumor-free tissues were obtained as described above and were incubated with FcBlock (clone 93, Biolegend) for 15min before staining with fluorescent conjugated antibodies for 45min at 4ºC. The cells were washed with staining buffer (PBS containing 0.5% BSA and 2 mM EDTA) and analyzed on a LSRII flow cytometer (BD). 7AAD was used to exclude dead cells and doublet cells were removed based on their forward/side scatter properties. Based on cell surface marker expression, H-SiglecF^low neutrophils (CD45⁺CD11b⁺Ly-6G⁺SiglecF^low) from healthy lung tissue and T-SiglecF^high neutrophils (CD45⁺CD11b⁺Ly-6G⁺SiglecF^high) as well as T-SiglecF^low neutrophils (CD45⁺CD11b⁺Ly-6G⁺SiglecF^low) from lung tumor tissue were identified by flow cytometry and analyzed for further marker expression identified by scRNASeq. Following Abs were purchased from BD: CD11b (557397, clone M1/70); Ly-6G (551461, 560599, clone 1A8); SiglecF (564514, clone E50–2440), Biolegend: CD45 (103126, clone 30-F11); CD73 (127205, clone TY/11.8) or R&D Systems: Clec5a (FAB1639P, clone 226402).

Zilionis R., Engblom C., Pfirschke C., Savova V., Zemmour D., Saatcioglu H.D., Krishnan I., Maroni G., Meyerovitz C.V., Kerwin C.M., Choi S., Richards W.G., Rienzo A.D., Tenen D.G., Bueno R., Levantini E., Pittet M.J, & Klein A.M. (2019). Single cell transcriptomics of human and mouse lung cancers reveals conserved myeloid populations across individuals and species. Immunity, 50(5), 1317-1334.e10.

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