Anti cd16 cd32 fc block 2 4g2

Manufactured by BD

Anti-CD16/CD32 Fc-Block (2,4G2) is a laboratory reagent that binds to the Fc receptors CD16 and CD32. This binding can block the non-specific interactions of Fc-containing molecules, such as antibodies, with these receptors.

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Lab products found in correlation

Facscanto 2 flow cytometer, by BD (1 mentions) Fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions) Ebio4b10, by Thermo Fisher Scientific (1 mentions) Foxp3 259d c7, by BD (1 mentions) Facscalibur, by BD (1 mentions) Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions) Brefeldin a, by Thermo Fisher Scientific (1 mentions) Ionomycin, by Merck Group (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) B220 ra3 6b2, by Thermo Fisher Scientific (1 mentions) Foxp3 fjk 16s, by Thermo Fisher Scientific (1 mentions) Cytofix cytoperm kit, by BD (1 mentions) Anti il 10 jes5 16e3, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Fc block (765 citations) Anti-CD16/CD32 (97 citations) Fc block (2.4G2, (41 citations) CD16/CD32 (41 citations) Fc block CD16/CD32 (39 citations) Anti-CD16/CD32 Fc block (34 citations) Anti-CD16/CD32 (2.4G2, (18 citations) CD16/CD32 (2.4G2, (16 citations) Anti-CD16/CD32 Fc block (clone 2.4G2; (7 citations) CD16/CD32 Fc block (2.4G2) (4 citations)

3 protocols using anti cd16 cd32 fc block 2 4g2

Comprehensive Flow Cytometry Staining Protocol

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For flow cytometric analysis, cell suspensions were pretreated with anti-CD16/CD32 Fc-Block (2,4G2; BD Biosciences) and stained by standard procedures. Dead cells were excluded by staining with Fixable Viability Dye eFluor780 (eBioscience). Cells were stained by using antibodies enlisted in Table 3.
Intracellular staining for flow cytometry was performed after surface staining by using the True-Nuclear Transcription Buffer Staining Set (BioLegend) according to the manufacturer’s instructions.
Cells were acquired at a FACSCantoII flow cytometer (BD Biosciences). If not indicated otherwise, numbers in the dot plots indicate the percentages of cells in the respective gates, numbers in the histograms indicate the mean fluorescence intensity (MFI).

Vadakumchery A., Faraidun H., Ayoubi O.E., Outaleb I., Schmid V., Abdelrasoul H., Amendt T., Khadour A., Setz C., Göhring K., Lodd K., Hitzing C., Alkhatib A., Bilal M., Benckendorff J., Al Shugri A.K., Brakebusch C.H., Engels N., Datta M., Hobeika E., Alsadeq A, & Jumaa H. (2022). The Small GTPase RHOA Links SLP65 Activation to PTEN Function in Pre B Cells and Is Essential for the Generation and Survival of Normal and Malignant B Cells. Frontiers in Immunology, 13, 842340.

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Murine Melanoma Immunotherapy Protocols

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The FBL cell line was a gift from Dr. Philip Greenberg (University of Washington) in 2008 and has been described previously (20 , 21 (link)). FBL has not been authenticated. The FBL cell line is maintained in vivo and cells are harvested from ascites fluid on the day of experiment setup. The HLA-A2⁺ human melanoma line MeWo was purchased from ATCC in 2014. Peptides from FBL-Gag (CCLCLTVFL) and ovalbumin (SIINFEKL) were obtained from GenScript. Mouse blocking antibodies to CTLA-4 (9D9), PD-1 (RMP1–14) and LAG-3 (C9B7W) were purchased from BioXCell. Human antibodies against CTLA-4, PD-1, and LAG-3 were provided by Bristol-Myers Squibb. All blocking antibodies were administered intraperitoneally (i.p.) at a dose of 100 µg/mouse every 3 days. Fluorochome-conjugated antibodies to mouse CD90.1 (OX-7), CD90.2 (53–2.1), IFNγ (XMG1.2), TNF (MP6-XT22), and anti-CD16/CD32 F_c block (2.4G2) and antibodies to human CD45 (HI30), CD3 (UCHT1), CD4 (RPA-T4), CD8 (SK1), and Foxp3 (259D/C7) were purchased from BD Biosciences. Fluorochrome-conjugated antibody to CD8 (53–6.7) was purchased from BioLegend. Fluorochrome-conjugated antibodies to mouse CD4 (GK1.5), NK1.1 (PK136), Eomes (Dan11mag), and Foxp3 (FJK-16s) and antibody to human T-bet (ebio4b10) were purchased from eBioscience.

Klevorn L.E., Berrien-Elliott M.M., Yuan J., Kuehm L.M., Felock G.D., Crowe S.A, & Teague R.M. (2016). Rescue of tolerant CD8+ T cells during cancer immunotherapy with IL2:antibody complexes. Cancer immunology research, 4(12), 1016-1026.

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Immune Cell Phenotyping by Flow Cytometry

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Single-cell suspensions were isolated from spleen, PeC and MLN tissues. For detection of intracellular IL-10 in B cells, isolated cells were resuspended and incubated for 5h with LPS (10 μg/ml, Sigma), phorbol 12-myristate 13-acetate (PMA, 50 ng/ml, Sigma), ionomycin (500 ng/ml, Sigma), and brefeldin A (3 μg/ml, eBioscience). To detect intracellular Foxp3 in T cells, the isolated cells were resuspended and exposed to the same agents, except LPS, for 4 h. Before staining for cell surface makers, cells were blocked with anti-CD16/CD32 Fc-block (2.4G2, BD Biosciences). Cells were fixed and permeabilized with Cytofix/Cytoperm kit (BD Biosciences) and then were stained with anti-IL-10 (JES5-16E3, eBioscience) and Foxp3 (FJK-16s, eBioscience). Abs against mouse surface proteins were: CD4 (GK1.5), CD5 (53-7.3), CD19 (eBio1D3), CD25 (PC61.5), and B220 (RA3-6B2), all from eBioscience Inc. (San Diego, CA). Cells were analyzed with FACScalibur (Becton Dickinson, San Jose, CA) and FlowJo version 10 software (TreeStar).

Kim A.R., Kim H.S., Kim D.K., Nam S.T., Kim H.W., Park Y.H., Lee D., Lee M.B., Lee J.H., Kim B., Beaven M.A., Kim H.S., Kim Y.M, & Choi W.S. (2016). Mesenteric IL-10-producing CD5+ regulatory B cells suppress cow’s milk casein-induced allergic responses in mice. Scientific Reports, 6, 19685.

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