Human fc block

Manufactured by BD

Sourced in United States, United Kingdom

The Human Fc block is a laboratory equipment product designed to facilitate protein purification and analysis. It serves as a tool for isolating and studying the Fc (fragment crystallizable) region of human immunoglobulins (antibodies). The Fc block provides a consistent and reliable method for researchers to capture and separate this specific protein domain from complex biological samples.

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Lab products found in correlation

Lsrfortessa, by BD (12 mentions) Facscanto 2, by BD (12 mentions) Cytoflex flow cytometer, by Beckman Coulter (7 mentions) Facsaria 3, by BD (6 mentions) Perm wash buffer, by BD (6 mentions) Facsdiva software, by BD (5 mentions) Cytofix, by BD (4 mentions) Facscanto, by BD (4 mentions) Live dead fixable aqua dead cell stain, by Thermo Fisher Scientific (4 mentions) Cytofix cytoperm fixation permeabilization solution kit, by BD (4 mentions) Lsrfortessa flow cytometer, by BD (4 mentions) Fortessa, by BD (4 mentions) Non enzymatic dissociation solution, by Corning (3 mentions) Cytofix cytoperm fixation permeabilization kit, by BD (3 mentions) Pd l1 bv421, by BD (3 mentions) Live dead 488 fixable green dead cell stain kit, by Thermo Fisher Scientific (3 mentions) Cytofix cytoperm solution, by BD (3 mentions) Fixable viability dye 780 apc cyanine 7, by Thermo Fisher Scientific (3 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (3 mentions) Live dead fixable near ir dead cell stain kit, by Thermo Fisher Scientific (3 mentions)

Close spelling variants of this product

NA/LE human BD Fc Block Murine and human Fc-Block Human Fc-receptor block Human BD Fc Block reagent Anti-mouse CD16/32 or human Fc block BD Pharmingen Human BD Fc Block Fc block

198 protocols using human fc block

Isolation and Characterization of CD206+ Alveolar Macrophages

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CD206+ AMs were enriched from donor BAL using the magnetic-based MACS® system (Miltenyi Biotech, Germany) and fluorescent activated cells sorting (FACS) as outlined previously 3, (link)20, (link)21 . Briefly, for MACS-based enrichment, BAL cells (1 × 10 7 ) were incubated with human Fc-block (BD Biosciences, USA) and human CD206 APC-Cy7 (clone 15-2, BioLegend, USA). CD206+ cells were enriched in MACS LS magnetic separation column and MidiMACS TM magnet. Cell counts were determined using a haemocytometer and trypan blue live/dead exclusion.
For FACS-based enrichment BAL cells were washed and incubated with nearinfrared fixable live/dead stain (Life Technologies Inc.) as per the manufacturer's instructions. After incubation with human Fc block (BD Pharmingen, Inc.), surface staining was performed with the following antibodies (fluorophore followed by clone in parentheses); CD45 (PE-Texas Red, H130), CD3 (FITC, OKT), TCR-β (BV421, IP26), CD206 (PercpCy5.5, 15.2). Cell sorting was carried out on Aria III (BD Biosciences) and AMs defined as live, CD45 + CD3 -TCR -CD206 + cells.
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint this version posted December 4, 2020. ; https://doi.org/10.1101/2020.12.04.410191 doi: bioRxiv preprint

McErlean P., Bell C.G., Hewitt R., Busharat Z., Ogger P.P., Ghai P., Albers G.J., Kingston S., Molyneaux P.L., Beck S., Lloyd C.M., Maher T.M., & Byrne A.J. (2020). Epigenetic alterations underlie airway macrophage differentiation and phenotype during lung fibrosis.

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Isolation and Characterization of Human Kidney Mononuclear Cells

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After informed consent, human kidney samples were collected from RCC patients undergoing nephrectomy surgery either before or after the clamping of renal pedicles. The kidney tissue was digested according to our protocol for isolation of KMNCs(36 ). Single-cell suspensions were pre-incubated with Human BD Fc Block (BD Biosciences) for 5 minutes and stained with monoclonal Ab anti-CD45 (HI30) BUV395 (BioLegend), TCRα/β (IP26) BV421, (BioLegend), CD4 (OKT4) FITC (eBioscience) for 30 minutes at 4 °C. Cells were fixed and permeabilized with fixation/permeabilization buffer (eBioscience) for 30 minutes at room temperature followed by another pre-incubation with Human BD Fc Block (BD Biosciences) for 5 minutes and intracellular staining with polyclonal Ab anti-NGAL APC (Assaypro, St. Charles, MO/Catalogue number: 31228–05161) at room temperature. APC-conjugated polyclonal Rabbit IgG was used as an isotype control (R&D systems, Minneapolis, MN) CD45⁺TCRα/β⁺CD4⁺ cells were analyzed for their intracellular NGAL expression using flow cytometry. The present study was conducted in accordance with the Declaration of Helsinki and approved by the Johns Hopkins Medicine Institutional Review Boards. No identifiable information was acquired during tissue collection.

Lee S.A., Noel S., Kurzhagen J.T., Sadasivam M., Pierorazio P.M., Arend L.J., Hamad A.R, & Rabb H. (2019). CD4+ T cell-derived NGAL modifies outcome from ischemic acute kidney injury. Journal of immunology (Baltimore, Md. : 1950), 204(3), 586-595.

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Quantifying Cell Surface ICAM-1 Expression

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Cells were harvested using non-enzymatic dissociation solution (Corning) and washed once with PBS. For staining of ICAM-1, cells were incubated in Human Fc Block (BD) for 10 min. Thereafter, cells were stained with FITC-conjugated antibody to ICAM-1 in the presence of Fc Block for 30 min at 4 °C. Cells were then washed 3× with PBS and analyzed by FACS Caliber (BD). Data analysis was performed using Flowjo V10 software (Ashland).

Park J., Hwang J.Y., Thore A., Kim S., Togano T., Hagiwara S., Park J.W, & Tse W. (2019). AF1q inhibited T cell attachment to breast cancer cell by attenuating Intracellular Adhesion Molecule-1 expression. Journal of cancer metastasis and treatment, 5, 17.

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ACE2 Expression on Immune Cells

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Whole blood (50 µL) was stained with LIVE/DEAD^® fixable cell stain kit (L/D-violet; Invitrogen) and then incubated (30 min) with human Fc block (BD Pharmingen, Franklin lakes, NJ, USA). ACE2 surface staining was conducted by Alexa Fluor^® 700-conjugated mouse anti-human ACE-2 (R&D Systems, Minneapolis, MN, USA) and APC-H7 mouse anti-human CD45 (BD Biosciences) antibodies. Data were acquired with a Gallios flow cytometer (Beckman Coulter, Pasadena, CA, USA) and analyzed using FlowJo V10 (TreeStar) or Kaluza 2 (Beckman Coulter) software. Circulating lymphocytes, monocytes and granulocytes were recognized by their distinct morphological features in forward and side scatter distribution.

Lye P., Dunk C.E., Zhang J., Wei Y., Nakpu J., Hamada H., Imperio G.E., Bloise E., Matthews S.G, & Lye S.J. (2021). ACE2 Is Expressed in Immune Cells That Infiltrate the Placenta in Infection-Associated Preterm Birth. Cells, 10(7), 1724.

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Flow Cytometry of Macrophage Subsets

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Macrophages were detached by incubation with ice-cold 5 mM EDTA in PBS for 15 min. Cells were washed once with FACS buffer (1% [wt/vol] BSA–0.5 mM EDTA in PBS), followed by an Fc receptor blockade with human Fc Block (BD Biosciences) in FACS buffer for 10 min at room temperature. Cells were stained for surface antigens with fluorochrome-conjugated (Brilliant Violet 421, FITC, phycoerythrin [PE]-Cy7, peridinin chlorophyll protein [PerCP]-Cy5.5, antigen-presenting cells [APC]) antibodies against human CD14 (MφP9), CD68 (Y1/82A), CD80 (L307.4), CD163 (GHI/61), and CD206 (19.2) (BD Biosciences) for 30 min at room temperature in the dark. Cells were washed once with FACS buffer before being fixed with 4% formaldehyde and analyzed by flow cytometry within 24 h. Samples were acquired using a BD LSRII flow cytometer equipped with BD FACS Diva software (BD Bioscience). FlowJo (TreeStar) was used for the final analysis.

Wagener J., MacCallum D.M., Brown G.D, & Gow N.A. (2017). Candida albicans Chitin Increases Arginase-1 Activity in Human Macrophages, with an Impact on Macrophage Antimicrobial Functions. mBio, 8(1), e01820-16.

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Assessing NK Cell Degranulation via CD107a

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A total of 18 healthy Chinese Southern Han individuals (11 of them with KIR AA genotype) were recruited from Shenzhen Blood Center. NK cells were isolated as above and incubated with K562 cells at a ratio of 5:1 for 5 h at 37°C. Optimal expression of CD107a on the cell surface of degranulating NK cells occurs between 4~6 h post stimulation (35 (link)). After the first 1 h, 4 μL monensin (BD Biosciences, CA, USA) was added to each 6 mL culture. The cells were then incubated with 2.5 uL human Fc block (BD Biosciences, CA, USA) at 25°C for 15 min, then stained with CD56, CD3, and CD107a mAbs. Antibodies used were: UCHT1 mouse anti-human CD3-FITC, H4A3 mouse anti-human CD107a-PECy5, and B159 mouse anti-human CD56-APC (all BD Biosciences, CA, USA). Flow cytometric analysis was performed using an ACCURI C6 machine (BD Biosciences, CA, USA) Surface expression of CD107a was assessed in CD56⁺CD3⁻ cells (36 (link)). To detect spontaneous degranulation, a control sample without target cells was included in every experiment and this information was used for background subtraction of the results.

Deng Z., Zhao J., Cai S., Qi Y., Yu Q., Martin M.P., Gao X., Chen R., Zhuo J., Zhen J., Zhang M., Zhang G., He L., Zou H., Lu L., Zhu W., Hong W., Carrington M, & Norman P.J. (2019). Natural Killer Cells Offer Differential Protection From Leukemia in Chinese Southern Han. Frontiers in Immunology, 10, 1646.

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Phenotypic Profiling of PBMCs and iPSC-Derived Cells

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Phenotypic analysis of PBMCs and human iPSC-derived cells was performed as described.22 24 27 (link) Briefly, single cell suspensions of PBMCs and human iPSC-derived cells were stained with a master mix of antibodies (Abs) for surface stains after Fc block (human Fc block; BD Biosciences) for 20 min at room temperature. Antibodies used in this study are listed in online supplemental table S2. Live/Dead Fixable Aqua Dead Cell Stain Kit (ThermoFisher Scientific) and LIVE/DEAD Fixable Near-IR Dead Cell Stain kit (ThermoFisher Scientific) were used to exclude dead cells from the analysis. Samples were analyzed using LSRII or LSRFortessa (BD Biosciences) with FlowJo software (TreeStar).

Makino K., Long M.D., Kajihara R., Matsueda S., Oba T., Kanehira K., Liu S, & Ito F. (2022). Generation of cDC-like cells from human induced pluripotent stem cells via Notch signaling. Journal for Immunotherapy of Cancer, 10(1), e003827.

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Profiling Immune Cell Subsets in Obesity

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Blood collection from human donors was approved by the Institutional Review Board of Rockefeller University, fully compliant with all relevant ethical regulations regarding research involving human participants, and obtained with informed consent. Fresh whole-blood samples were obtained from healthy female donors (including 5 lean donors, and 2 obese donors with BMI ≥ 35; donors were all postmenopausal) at Rockefeller University. Red blood cells were lysed directly (Pharm Lyse, BD Biosciences), cells were counted, incubated with human Fc block (1 h; BD Biosciences; 1:100 10⁻⁶ cells), incubated with conjugated antibodies (1 h), and stained with DAPI (10 min). Antibodies and dilutions are listed in Supplementary Table 3. Cell surface markers were used to define different cell populations as follows: peripheral blood neutrophils (CD45⁺CD11b⁺CD66b⁺CD16⁺CD14^lo), eosinophils (CD45⁺CD11b⁺CD66b⁺CD16⁻CD14^lo), non-classical monocytes (CD45⁺CD11b⁺CD66b⁻CD16⁺CD14^lo), intermediate monocytes (CD45⁺CD11b⁺CD66b⁻CD16⁺CD14^hi) and classical monocytes (CD45⁺CD11b⁺CD66b⁻CD16⁻CD14⁺). An antibody targeting IL5Rα was also included for all staining. Eosinophils were used as a positive gating control for IL5Rα positivity as this is a canonical marker/signalling pathway for this cell type^{53 (link)}.

Quail D.F., Olson O.C., Bhardwaj P., Walsh L.A., Akkari L., Quick M.L., Chen I.C., Wendel N., Ben-Chetrit N., Walker J., Holt P.R., Dannenberg A.J, & Joyce J.A. (2017). Obesity alters the lung myeloid cell landscape to enhance breast cancer metastasis through IL5 and GM-CSF. Nature cell biology, 19(8), 974-987.

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Detecting CD68 in Leukemia Cells

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Leukaemia cell lines were blocked in FACS buffer (PBS + 0.5% FBS) containing human Fc block (BD Biosciences; 564220) for 20 min at room temperature. Cells were fixed, permeabilized using the BD Cytofix/Cytoperm fixation/permeabilization kit (BD Bioscience) and later stained with anti-human CD68-PECy7 (BioLegend; 333816). Sample acquisition was performed on a BD LSR II flow cytometer and the mean fluorescence intensities (MFI) were calculated using FlowJo 9.3 software for Mac OS X (TreeStar).

Trujillo-Alonso V., Pratt E.C., Zong H., Lara-Martinez A., Kaittanis C., Rabie M.O., Longo V., Becker M.W., Roboz G.J., Grimm J, & Guzman M.L. (2019). FDA-approved ferumoxytol displays anti-leukaemia efficacy against cells with low ferroportin levels. Nature nanotechnology, 14(6), 616-622.

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Phenotypic Profiling of Immortalized Lung Pericytes

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Immortalized human lung pericytes at passage 16 were detached with Accutase, washed and suspended in Dulbecco’s phosphate-buffered saline containing 2% BSA and 0.75 mM NaN₃. After blocking CD16/CD32 with Human Fc Block (BD Biosciences, Franklin Lakes, NJ), cells were stained with the following antibodies (Biolegend, San Diego, CA): APC-anti human CD34, APC-anti human CD45, PE-anti human CD31, APC-anti human CD44, PE-anti human CD146, PE-anti human CD90, PE-anti human CD73 and PE-anti human CD140b, and isotype controls diluted according to the manufacturer’s instructions. Cells were analyzed using a CytoFLEX flow cytometer and CytExpert 2.3 software (Beckman Coulter, Brea, CA).

Li P., Wu Y., Goodwin A.J., Halushka P.V., Wilson C.L., Schnapp L.M, & Fan H. (2021). Generation of a new immortalized human lung pericyte cell line: a promising tool for human lung pericyte studies. Laboratory investigation; a journal of technical methods and pathology, 101(5), 625-635.

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