Fcr binding inhibitor

Manufactured by Thermo Fisher Scientific

The FcR-binding inhibitor is a laboratory equipment product designed to inhibit the binding of Fc receptors (FcRs) to their ligands. It is a useful tool for researchers studying immune cell signaling and antibody-mediated functions.

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Lab products found in correlation

Efluor 450, by Thermo Fisher Scientific (2 mentions) Fixation permeabilization solution kit, by Thermo Fisher Scientific (2 mentions) Macsquant analyser 10, by Miltenyi Biotec (2 mentions) Facscanto 2 flow cytometer, by BD (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Attune nxt, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Human FcR binding inhibitor (11 citations) Hu FcR Binding Inhibitor (2 citations)

4 protocols using fcr binding inhibitor

Multiparametric analysis of cryopreserved PBMCs

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Cryopreserved peripheral blood mononuclear cells (PBMC) were thawed and stained with fixable viability dye eFluor 450 (eBioscience) for 30 min at 4^oC. FcR-binding inhibitor (eBioscience) was added to all cells followed by incubation with antibodies targeting surface markers (see Life Sciences Reporting Summary for list of antibodies and dilutions used).
Prior to addition of antibodies targeting intracellular targets, cells were permeabilized and fixed using Fixation/Permeabilization Solution Kit (eBioscience). Samples were acquired on a MACSquant Analyser 10 (Miltenyi Biotec) and data was analyzed using the FlowJo software (Treestar). The gating strategy is illustrated in Supplementary Fig. 6.

Formenti S.C., Rudqvist N.P., Golden E., Cooper B., Wennerberg E., Lhuillier C., Vanpouille-Box C., Friedman K., de Andrade L.F., Wucherpfennig K.W., Heguy A., Imai N., Gnjatic S., Emerson R.O., Zhou X.K., Zhang T., Chachoua A, & Demaria S. (2018). Radiotherapy induces responses of lung cancer to CTLA-4 blockade. Nature medicine, 24(12), 1845-1851.

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Multiparametric analysis of cryopreserved PBMCs

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Cytokine Profiling of Stimulated PBMCs

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Heparinized WB was collected and processed within 2 hours. PBMC were isolated by standard methods on Ficoll-Paque Plus (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) and incubated with stimuli at 37°C. Brefeldin at 10ug/mL (Sigma) was added after 1 h or 20 h of stimulation. ICS was performed after 24 h of incubation. Unstimulated cells were used as a negative control. PBMC were stained for vitality and then fixed in 2% paraformaldehyde. Therefore, the cells were resuspended in the PBS-2% FCS-0.5% saponin-2mM EDTA-1% FcR- binding inhibitor (eBioscience) buffer and stained with mAbs for surface markers and intracellular cytokines. At least 300,000 events were acquired using a FACSCanto II flow cytometer (BD Biosciences).

Petrone L., Vanini V., Petruccioli E., Ettorre G.M., Schininà V., Busi Rizzi E., Ludovisi A., Corpolongo A., Ippolito G., Pozio E., Teggi A, & Goletti D. (2015). Polyfunctional Specific Response to Echinococcus Granulosus Associates to the Biological Activity of the Cysts. PLoS Neglected Tropical Diseases, 9(11), e0004209.

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Activation-dependent CD71 expression

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Jurkat cells and primary CD4⁺ T cells from the activation procedure were collected at different days post activation, washed with PBS and resuspended in 100 μl of PBS. Per sample, 10 μl of 1:10 diluted FcR binding inhibitor (eBioscience, Frankfurt, Germany) in PBS was added, mixed and incubated for 5 min at 4 °C. Subsequently, 1 μl of undiluted anti-CD71 antibody (eBioscience) was added to appropriate samples, while isotype controls were stained with IgG1 antibody (eBioscience) and blank samples were left unstained. All samples were vortexed and incubated for another 30 min at 4 °C protected from light. After washing with PBS 3 times, cells were resuspended in 400 μl PBS/2 mM EDTA and sample were analyzed using an Attune ® NxT flow cytometer (Thermo Fisher Scientific) with 488 excitation and 574/26 emission filter. The cells of all samples were gated according to morphology based on forward/sideward scattering, and 10.000 events were evaluated per sample.

Kandil R., Baldassi D., Böhlen S., Müller J.T., Keul D.C., Bargmann T., Dehmel S., Xie Y., Mehta A., Sewald K, & Merkel O.M. (2023). Targeted GATA3 knockdown in activated T Cells via pulmonary siRNA delivery as novel therapy for allergic asthma. Journal of controlled release : official journal of the Controlled Release Society, 354, 305-315.

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