Fc blocking antibody

Manufactured by BioLegend

Sourced in United States

The Fc blocking antibody is a laboratory reagent used to block non-specific binding of antibodies to Fc receptors on cells. It helps to reduce background signal and improve the specificity of immunoassays and flow cytometry experiments.

Automatically generated - may contain errors

Lab products found in correlation

Facsaria 2, by BD (2 mentions) Collagenase 2, by Merck Group (2 mentions) Dulbecco modified eagle medium (dmem), by Bio-Rad (2 mentions) Dnase, by Bio-Rad (2 mentions) Apc conjugated isotype controls, by BioLegend (2 mentions) Apc conjugated anti f4 80, by BioLegend (2 mentions) Anti cd140b, by BioLegend (2 mentions) Facscelesta, by BD (1 mentions) Collagenase type 4, by Thermo Fisher Scientific (1 mentions) Facscanto 2 cytometer, by BD (1 mentions) Live dead fixable dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Rbc lysis buffer, by Thermo Fisher Scientific (1 mentions) Foxp3 transcription factor staining buffer, by Thermo Fisher Scientific (1 mentions) Deoxyribonuclease 1, by Roche (1 mentions) Facs fortessa, by BD (1 mentions) 7 aminoactinomycin d, by BioLegend (1 mentions) Cd11b clone m1 70, by Thermo Fisher Scientific (1 mentions) Lsrfortessa, by BD (1 mentions) Zombie uv viability dye, by BioLegend (1 mentions)

Spelling variants of this product

Fc receptor blocking antibody (7 citations) Fc blocking antibody against CD16/CD32 (3 citations) CD16/32 Fc-blocking antibody (2 citations) Fc blocking antibody (Clone 93, (2 citations) CD16/32 Fc‐receptor blocking antibody (2 citations) Anti-Fc receptor-blocking antibody (anti-CD16/CD32; (2 citations)

7 protocols using fc blocking antibody

Isolation of Kidney Stromal Cells

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

For tissue preparation, the obstructed kidney was minced and incubated with 2mg/mL collagenase II (Sigma), 1mL DMEM, and DNAse (BioRad 10ul/mL) in a shaker at 37C for 1 hour. The tissue was filtered (70 μm), centrifuged, and incubated in red blood cell lysis buffer (15.5mM NH₄Cl, 1mM KHCO₃, 10 μM EDTA) at 37C for 5 minutes, neutralized with PBS, centrifuged, and resuspended in PBS with 1%FBS. The tissue was passed through a 50 μM strainer, and tissue from COL-Cre mice (and controls) was passed through a 24 μM filter to remove the glomeruli. The FACSAria II from BD Biosciences in the Research Flow Cytometry Core Laboratory at the Nashville VA Medical Center was used for sorting of GFP+ cells.
For flow cytometry, samples were incubated with Fc blocking antibody (Biolegend) on ice, then incubated with APC-conjugated anti-F4/80, anti-CD140b (PDGFRβ), or the appropriate APC-conjugated isotype controls (Biolegend) for 30 minutes at room temperature. Cells were analyzed on a FACS Canto II cytometer with FACSDiVa v6.1 software for data acquisition and analysis at the Nashville VA Medical Center’s Flow Cytometry Laboratory.

Neelisetty S., Alford C., Reynolds K., Woodbury L., Nlandu-khodo S., Yang H., Fogo A.B., Hao C.M., Harris R.C., Zent R, & Gewin L. (2015). Renal fibrosis is not reduced by blocking transforming growth factor-β signaling in matrix-producing interstitial cells. Kidney international, 88(3), 503-514.

+ Open protocol

+ Expand

Lung Cell Isolation and Flow Cytometry

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

BALF or lung single cell suspension was obtained after PBS, LPS or heated-P. aeruginosa i.n. instillation. Lung cells were collected by centrifugation and then incubated with Fc blocking antibody (purified anti-mouse CD16/32, BioLegend) on ice for 15 min. After the cells were stained with fluorescence-conjugated antibodies for 30 min on ice, the cells were washed and subjected to FACS (BD, FACSCelesta). The FACS data were analyzed using Flow Jo software (TreeStar,Inc). All antibodies used for flow cytometry were anti-mouse antigens.

Cheng X., Jiang W., Chen Y., Zou B., Wang Z., Gan L., Xiao Z., Li C., Yu C.Y., Lu Y., Han Z., Zeng J., Gu J., Chu T., Fu M., Chu Y., Zhang W., Tang J, & Lu M. (2023). Acyloxyacyl hydrolase promotes pulmonary defense by preventing alveolar macrophage tolerance. PLOS Pathogens, 19(7), e1011556.

+ Open protocol

+ Expand

Tumor Dissociation and Flow Cytometry

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

Tumor-bearing mice were euthanized, and resected tumors were shredded into small pieces and incubated in collagenase-containing buffer: collagenase type IV (100 U/ml; Invitrogen), deoxyribonuclease I (50 μg/ml; Roche), and 10% FBS in RPMI 1640 medium for 45 min. After incubation, cells were treated with RBC lysis buffer (Thermo Fisher Scientific) and passed through a 100-μm cell strainer to remove debris. The cell pellet was dissolved by 2% FBS in PBS and used for flow cytometry analysis. Isolated cells were initially stained with the LIVE/DEAD Fixable Dead Cell Stain Kit (Invitrogen). Cells were subsequently incubated with Fc-blocking antibody (BioLegend) and then stained with monoclonal antibodies for several surface and intracellular antigens as listed in Supplementary Methods. Foxp3/Transcription Factor Staining Buffer (eBioscience) was used for intracellular staining. Sample acquisition was performed on a BD FACSCanto II cytometer (BD Biosciences) equipped with Diva software and analyzed using FlowJo software (BD Biosciences). Antibody clone numbers for immune analysis are listed in the Supplementary Materials.

Naito Y., Koyama S., Masuhiro K., Hirai T., Uenami T., Inoue T., Osa A., Machiyama H., Watanabe G., Sax N., Villa J., Kinugasa-Katayama Y., Nojima S., Yaga M., Hosono Y., Okuzaki D., Satoh S., Tsuda T., Nakanishi Y., Suga Y., Morita T., Fukushima K., Nishide M., Shiroyama T., Miyake K., Iwahori K., Hirata H., Nagatomo I., Yano Y., Tamiya M., Kumagai T., Takemoto N., Inohara H., Yamasaki S., Yamashita K., Aoshi T., Akbay E.A., Hosen N., Shintani Y., Takamatsu H., Mori M., Takeda Y, & Kumanogoh A. (2023). Tumor-derived semaphorin 4A improves PD-1–blocking antibody efficacy by enhancing CD8+ T cell cytotoxicity and proliferation. Science Advances, 9(20), eade0718.

+ Open protocol

+ Expand

Immunophenotyping of Cell Samples

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

Cells were incubated with unlabeled isotype control antibody and Fc blocking antibody for 15 min (BioLegend, New Jersey, USA). After that, fluorescently coupled antibodies were added, and the cells were incubated on ice for 30 min. After washing with PBS, cells were analyzed on FACS Fortessa (Becton Dickinson, Franklin Lakes, New Jersey, USA) using FlowJo V.10.1 software (Tree Star, California, USA). Cell debris and dead cells were excluded by forward and side scatter gating and 7-amino-actinomycin D (BioLegend) staining.

Zhang Y., Huo F., Cao Q., Jia R., Huang Q., Wang Z.A., Theodorescu D., Lv Q., Li P, & Yan C. (2022). FimH confers mannose-targeting ability to Bacillus Calmette-Guerin for improved immunotherapy in bladder cancer. Journal for Immunotherapy of Cancer, 10(3), e003939.

+ Open protocol

+ Expand

Isolation of Kidney Stromal Cells

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

+ Open protocol

+ Expand

Quantifying Murine MDSC in Bone Marrow

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

Multicolor flow cytometry (FCM) was performed to observe the percentage of MDSCs in the bone marrow of CRC mice. After being freed of muscles and tendons, the femurs and tibiae of mice were placed in 70% ethanol for 2 min and subsequently washed in PBS, then a syringe was used to flush bone marrow cells from the femurs and tibias with PBS. After red blood cells were lysed, flushing fluid was filtered through a 100-μm membrane to obtain suspension of single cells. Cells were incubated with Fc blocking antibody (BioLegend, 101319) for 15 min and then stained with fluorescence-conjugated antibodies of surface markers CD11b (clone M1/70, eBioscience, 11-0112-82) and Gr-1 (clone RB6-8C5, Biogems, 83122-80-25) for 30 min. The samples were detected by a BD FACS Aria Fusion Flow Cytometry Cell Sorter (BD Biosciences), and the data were analyzed using FlowJo v.9 software (FlowJo LLC).

Gu J., Lv X., Li W., Li G., He X., Zhang Y., Shi L, & Zhang X. (2023). Deciphering the mechanism of Peptostreptococcus anaerobius-induced chemoresistance in colorectal cancer: the important roles of MDSC recruitment and EMT activation. Frontiers in Immunology, 14, 1230681.

+ Open protocol

+ Expand

Comprehensive Immune Profiling of Tumor Cells

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

Following tumor dissociation cells were stained with ZombieUV viability dye (Biolegend), before being resuspended in PBS + 1% BSA. Approximately 1x10⁶ cells were aliquoted in 5 ml tubes and incubated with Fc Blocking antibody (Biolegend). Antibodies used for immune profiling are detailed in Supplementary file 1, and master mixes were prepared before incubation with cells. After washing, cells were fixed and permeabilised overnight using FoxP3/transcription factor staining buffer set following manufacturer’s instructions (eBiosceince, ThermoFischer), before staining with intracellular antibody master mixes. Finally, cells were washed before being acquired on BD LSR Fortessa (BD Biosciences). Gating and analysis of flow cytometry data was conducted using FlowJo (Version 10.8, Tree Star). Gating of major populations are demonstrated in Figure 2—figure supplement 1 and Figure 3—figure supplement 1. For immunophenotyping experiments, sample size was chosen to ensure biological differences could be determined, with minimum mice numbers to allow for collection and processing in one day. Samples were excluded if intracellular staining was unsuccessful.

Webb E.R., Dodd G.L., Noskova M., Bullock E., Muir M., Frame M.C., Serrels A, & Brunton V.G. (2023). Kindlin-1 regulates IL-6 secretion and modulates the immune environment in breast cancer models. eLife, 12, e85739.

+ 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!