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fc block buffer  (Bio-Rad)


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    Bio-Rad fc block buffer
    Fc Block Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd16/Rat+anti+Mouse+CD16%2FCD32/pm41663544-252-16-18
    Average 93 stars, based on 35 article reviews
    fc block buffer - by Bioz Stars, 2026-10
    93/100 stars

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    Related Articles

    Flow Cytometry:

    Article Title: A novel real time imaging platform to quantify macrophage phagocytosis.
    Article Snippet: Red blood cells were lysed with BD Pharm LyseTM (BD Biosciences, Oxford, UK) as instructed by the manufacturer and were re-suspended in 0.5 ml PBS/1% FBS for further use. .. Lung and peritoneal harvested cells were blocked with 10 lg/ml rat anti-mouse CD16/CD32 (AbD Serotec, Oxford, UK) and 1.17 mg/ ml murine immunoglobulin G (IgG) (Jackson ImmunoResearch Laboratories, PA, USA) in Flow Cytometry buffer (PBS; 2% FBS, 25 mM HEPES, 5 mM EDTA) for 15 min on ice. .. Cells were stained with anti-CD45 (30-F11; BD Pharmigen), anti-CD11c (HL3; BD Pharmigen), anti-Siglec F (E50-2440; BD Pharmigen), anti-CD11b (M1/70; Biolegend, London, UK) and anti-F4/80 (CI:A3-1; Bio-rad) at 2 lg/ml in Flow Cytometry buffer for 30 min on ice protected from light.

    Article Title: A novel real time imaging platform to quantify macrophage phagocytosis
    Article Snippet: Red blood cells were lysed with BD Pharm LyseTM (BD Biosciences, Oxford, UK) as instructed by the manufacturer and were re-suspended in 0.5 ml PBS/1% FBS for further use. .. Lung and peritoneal harvested cells were blocked with 10 μg/ml rat anti-mouse CD16/CD32 (AbD Serotec, Oxford, UK) and 1.17 mg/ml murine immunoglobulin G (IgG) (Jackson ImmunoResearch Laboratories, PA, USA) in Flow Cytometry buffer (PBS; 2% FBS, 25 mM HEPES, 5 mM EDTA) for 15 min on ice. .. Cells were stained with anti-CD45 (30-F11; BD Pharmigen), anti-CD11c (HL3; BD Pharmigen), anti-Siglec F (E50-2440; BD Pharmigen), anti-CD11b (M1/70; Biolegend, London, UK) and anti-F4/80 (CI:A3-1; Bio-rad) at 2 μg/ml in Flow Cytometry buffer for 30 min on ice protected from light.

    Incubation:

    Article Title: A broadly adaptable protocol for isolating Kupffer cells from non-model species: application to Mastomys natalensis and its susceptibility to Old World mammarenaviruses
    Article Snippet: .. For flow cytometric analysis, the cells were harvested and first incubated with Fc-block buffer (BioRad #MCA2305) for 15 mins. .. After that, the cells were incubated with antibodies against CD11b (Biolegend #101216, 1:200), MHC-II (Invitrogen #12-5980-82, 1:200), CD80 (Biolegend #104713, 1:200), Iba1 (Fujifilm #019-19741, 1:50) and ZombieNIR (Biolegend #423106, 1:200) for 15 min at room temperature in the dark.

    Article Title: Lassa virus circumvents macrophage and dendritic cell antiviral defences in its natural reservoir, the Natal multimammate mouse (Mastomys natalensis).
    Article Snippet: .. For flow cytometric analysis, the cells were harvested at the corresponding timepoints and first incubated with Fc-block buffer (BioRad #MCA2305) for 15min. .. In parallel, the cells were stimulated with8 μg/mLofLPS (InvivoGen), 25 ng/mLofPoly I:C (HMW, InvivoGen) or 30 HAU of SeV, for the corresponding timepoints.

    other:

    Article Title: Lassa virus circumvents macrophage and dendritic cell antiviral defences in its natural reservoir, the Natal multimammate mouse ( Mastomys natalensis )
    Article Snippet: Lastly, the samples were stained using the TBM buffer (RecomBlot, Mikrogen) following the manufacturer’s instructions.

    Control:

    Article Title: CD13/aminopeptidase N is a negative regulator of mast cell activation.
    Article Snippet: Antigen-inducedmast cell (MC) activation via cross-linking of IgE-bound high-affinity receptors for IgE (Fc«RI) underlies type I allergy and anaphylactic shock.. Comprehensive knowledge of Fc«RI regulation is thus required.. We have identified a functional interaction between Fc«RI and CD13 in murineMCs.

    Purification:

    Article Title: Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Allophycocyanin/cyanine 7 anti-mouse CD45 (1:100) BioLegend Cat#103116 BV605 anti-mouse CD31 (1:100) BioLegend Cat#102427 (Continued on next page) 2 STAR Protocols 4, 102480, September 15, 2023 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Purified Rat Anti-Mouse CD16/CD32 Clone 2.4G2 (RUO) (1:100) BD Pharmingen Cat #553142 Chemicals, peptides, and recombinant proteins 103 Tris buffered saline (TBS) Bio-Rad Cat#1706463 16% Paraformaldehyde (formaldehyde) aqueous solution, EM grade Thomas Scientific Cat# C993M26 Ammonium chloride (NH4Cl), crystal VWR Chemicals Cat#12125-02-9 Fetal bovine serum (FBS) Gibco Cat#10437-028 DAPI (1:100) BioLegend Cat#422801 PBS (103), pH 7.4 Thermo Fisher Scientific Cat#70011069 PBS (13), pH 7.4 Thermo Fisher Scientific Cat#10010023 Percoll Sigma-Aldrich Cat#17-0891-02 Prolong Gold antifade reagent with DAPI Invitrogen Cat#P36980 RPMI 1640 Medium Thermo Fisher Scientific Cat#11875093 Sodium azide, 5% Fisher Scientific Cat#71448-16 Triton X-100 American Bio Cat#9002-93-1 UltraPure 0.5 M EDTA, pH 8.0 Life Technologies Cat#15575020 Other Cover Glass Thickness 1 1/2 22 3 22 mm Corning Cat#2850-22 Epredia Single Cytofunnel with White Filter Cards Thermo Fisher Scientific Cat#5991040 Epredia Cytoclip Stainless-Steel Slide Clip Thermo Fisher Scientific Cat#59910052 Glass Coplin Jar Thermo Fisher Scientific Cat#E94 Unifrost Plus Microscope Slides Adhesive, Treated Surfaces Azer Scientific Cat#EMS200G+ ImmEdge Hydrophobic Barrier PAP Pen Vector Laboratories Cat#H-4000 Thermo Scientific Cytospin 4 Cytocentrifuge Thermo Fisher Scientific Cat# A78300003 Sorvall Legend T Benchtop Centrifuge Rotor: 75006445 Sorvall Legend Cat#SO-LEGT BD FACS Aria II sorter BD Biosciences N/A Beckman Coulter Allegra 6 Centrifuge Rotor: GH-3.8 Swing Bucket Rotor Beckman Coulter Cat#BE-A6 Leica TCS SP8 Leica N/A Software and algorithms FlowJo 10 BD http://flowjo.com ll OPEN ACCESSProtocol ..

    Recombinant:

    Article Title: Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Allophycocyanin/cyanine 7 anti-mouse CD45 (1:100) BioLegend Cat#103116 BV605 anti-mouse CD31 (1:100) BioLegend Cat#102427 (Continued on next page) 2 STAR Protocols 4, 102480, September 15, 2023 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Purified Rat Anti-Mouse CD16/CD32 Clone 2.4G2 (RUO) (1:100) BD Pharmingen Cat #553142 Chemicals, peptides, and recombinant proteins 103 Tris buffered saline (TBS) Bio-Rad Cat#1706463 16% Paraformaldehyde (formaldehyde) aqueous solution, EM grade Thomas Scientific Cat# C993M26 Ammonium chloride (NH4Cl), crystal VWR Chemicals Cat#12125-02-9 Fetal bovine serum (FBS) Gibco Cat#10437-028 DAPI (1:100) BioLegend Cat#422801 PBS (103), pH 7.4 Thermo Fisher Scientific Cat#70011069 PBS (13), pH 7.4 Thermo Fisher Scientific Cat#10010023 Percoll Sigma-Aldrich Cat#17-0891-02 Prolong Gold antifade reagent with DAPI Invitrogen Cat#P36980 RPMI 1640 Medium Thermo Fisher Scientific Cat#11875093 Sodium azide, 5% Fisher Scientific Cat#71448-16 Triton X-100 American Bio Cat#9002-93-1 UltraPure 0.5 M EDTA, pH 8.0 Life Technologies Cat#15575020 Other Cover Glass Thickness 1 1/2 22 3 22 mm Corning Cat#2850-22 Epredia Single Cytofunnel with White Filter Cards Thermo Fisher Scientific Cat#5991040 Epredia Cytoclip Stainless-Steel Slide Clip Thermo Fisher Scientific Cat#59910052 Glass Coplin Jar Thermo Fisher Scientific Cat#E94 Unifrost Plus Microscope Slides Adhesive, Treated Surfaces Azer Scientific Cat#EMS200G+ ImmEdge Hydrophobic Barrier PAP Pen Vector Laboratories Cat#H-4000 Thermo Scientific Cytospin 4 Cytocentrifuge Thermo Fisher Scientific Cat# A78300003 Sorvall Legend T Benchtop Centrifuge Rotor: 75006445 Sorvall Legend Cat#SO-LEGT BD FACS Aria II sorter BD Biosciences N/A Beckman Coulter Allegra 6 Centrifuge Rotor: GH-3.8 Swing Bucket Rotor Beckman Coulter Cat#BE-A6 Leica TCS SP8 Leica N/A Software and algorithms FlowJo 10 BD http://flowjo.com ll OPEN ACCESSProtocol ..

    Saline:

    Article Title: Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Allophycocyanin/cyanine 7 anti-mouse CD45 (1:100) BioLegend Cat#103116 BV605 anti-mouse CD31 (1:100) BioLegend Cat#102427 (Continued on next page) 2 STAR Protocols 4, 102480, September 15, 2023 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Purified Rat Anti-Mouse CD16/CD32 Clone 2.4G2 (RUO) (1:100) BD Pharmingen Cat #553142 Chemicals, peptides, and recombinant proteins 103 Tris buffered saline (TBS) Bio-Rad Cat#1706463 16% Paraformaldehyde (formaldehyde) aqueous solution, EM grade Thomas Scientific Cat# C993M26 Ammonium chloride (NH4Cl), crystal VWR Chemicals Cat#12125-02-9 Fetal bovine serum (FBS) Gibco Cat#10437-028 DAPI (1:100) BioLegend Cat#422801 PBS (103), pH 7.4 Thermo Fisher Scientific Cat#70011069 PBS (13), pH 7.4 Thermo Fisher Scientific Cat#10010023 Percoll Sigma-Aldrich Cat#17-0891-02 Prolong Gold antifade reagent with DAPI Invitrogen Cat#P36980 RPMI 1640 Medium Thermo Fisher Scientific Cat#11875093 Sodium azide, 5% Fisher Scientific Cat#71448-16 Triton X-100 American Bio Cat#9002-93-1 UltraPure 0.5 M EDTA, pH 8.0 Life Technologies Cat#15575020 Other Cover Glass Thickness 1 1/2 22 3 22 mm Corning Cat#2850-22 Epredia Single Cytofunnel with White Filter Cards Thermo Fisher Scientific Cat#5991040 Epredia Cytoclip Stainless-Steel Slide Clip Thermo Fisher Scientific Cat#59910052 Glass Coplin Jar Thermo Fisher Scientific Cat#E94 Unifrost Plus Microscope Slides Adhesive, Treated Surfaces Azer Scientific Cat#EMS200G+ ImmEdge Hydrophobic Barrier PAP Pen Vector Laboratories Cat#H-4000 Thermo Scientific Cytospin 4 Cytocentrifuge Thermo Fisher Scientific Cat# A78300003 Sorvall Legend T Benchtop Centrifuge Rotor: 75006445 Sorvall Legend Cat#SO-LEGT BD FACS Aria II sorter BD Biosciences N/A Beckman Coulter Allegra 6 Centrifuge Rotor: GH-3.8 Swing Bucket Rotor Beckman Coulter Cat#BE-A6 Leica TCS SP8 Leica N/A Software and algorithms FlowJo 10 BD http://flowjo.com ll OPEN ACCESSProtocol ..

    Microscopy:

    Article Title: Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Allophycocyanin/cyanine 7 anti-mouse CD45 (1:100) BioLegend Cat#103116 BV605 anti-mouse CD31 (1:100) BioLegend Cat#102427 (Continued on next page) 2 STAR Protocols 4, 102480, September 15, 2023 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Purified Rat Anti-Mouse CD16/CD32 Clone 2.4G2 (RUO) (1:100) BD Pharmingen Cat #553142 Chemicals, peptides, and recombinant proteins 103 Tris buffered saline (TBS) Bio-Rad Cat#1706463 16% Paraformaldehyde (formaldehyde) aqueous solution, EM grade Thomas Scientific Cat# C993M26 Ammonium chloride (NH4Cl), crystal VWR Chemicals Cat#12125-02-9 Fetal bovine serum (FBS) Gibco Cat#10437-028 DAPI (1:100) BioLegend Cat#422801 PBS (103), pH 7.4 Thermo Fisher Scientific Cat#70011069 PBS (13), pH 7.4 Thermo Fisher Scientific Cat#10010023 Percoll Sigma-Aldrich Cat#17-0891-02 Prolong Gold antifade reagent with DAPI Invitrogen Cat#P36980 RPMI 1640 Medium Thermo Fisher Scientific Cat#11875093 Sodium azide, 5% Fisher Scientific Cat#71448-16 Triton X-100 American Bio Cat#9002-93-1 UltraPure 0.5 M EDTA, pH 8.0 Life Technologies Cat#15575020 Other Cover Glass Thickness 1 1/2 22 3 22 mm Corning Cat#2850-22 Epredia Single Cytofunnel with White Filter Cards Thermo Fisher Scientific Cat#5991040 Epredia Cytoclip Stainless-Steel Slide Clip Thermo Fisher Scientific Cat#59910052 Glass Coplin Jar Thermo Fisher Scientific Cat#E94 Unifrost Plus Microscope Slides Adhesive, Treated Surfaces Azer Scientific Cat#EMS200G+ ImmEdge Hydrophobic Barrier PAP Pen Vector Laboratories Cat#H-4000 Thermo Scientific Cytospin 4 Cytocentrifuge Thermo Fisher Scientific Cat# A78300003 Sorvall Legend T Benchtop Centrifuge Rotor: 75006445 Sorvall Legend Cat#SO-LEGT BD FACS Aria II sorter BD Biosciences N/A Beckman Coulter Allegra 6 Centrifuge Rotor: GH-3.8 Swing Bucket Rotor Beckman Coulter Cat#BE-A6 Leica TCS SP8 Leica N/A Software and algorithms FlowJo 10 BD http://flowjo.com ll OPEN ACCESSProtocol ..

    Adhesive:

    Article Title: Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Allophycocyanin/cyanine 7 anti-mouse CD45 (1:100) BioLegend Cat#103116 BV605 anti-mouse CD31 (1:100) BioLegend Cat#102427 (Continued on next page) 2 STAR Protocols 4, 102480, September 15, 2023 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Purified Rat Anti-Mouse CD16/CD32 Clone 2.4G2 (RUO) (1:100) BD Pharmingen Cat #553142 Chemicals, peptides, and recombinant proteins 103 Tris buffered saline (TBS) Bio-Rad Cat#1706463 16% Paraformaldehyde (formaldehyde) aqueous solution, EM grade Thomas Scientific Cat# C993M26 Ammonium chloride (NH4Cl), crystal VWR Chemicals Cat#12125-02-9 Fetal bovine serum (FBS) Gibco Cat#10437-028 DAPI (1:100) BioLegend Cat#422801 PBS (103), pH 7.4 Thermo Fisher Scientific Cat#70011069 PBS (13), pH 7.4 Thermo Fisher Scientific Cat#10010023 Percoll Sigma-Aldrich Cat#17-0891-02 Prolong Gold antifade reagent with DAPI Invitrogen Cat#P36980 RPMI 1640 Medium Thermo Fisher Scientific Cat#11875093 Sodium azide, 5% Fisher Scientific Cat#71448-16 Triton X-100 American Bio Cat#9002-93-1 UltraPure 0.5 M EDTA, pH 8.0 Life Technologies Cat#15575020 Other Cover Glass Thickness 1 1/2 22 3 22 mm Corning Cat#2850-22 Epredia Single Cytofunnel with White Filter Cards Thermo Fisher Scientific Cat#5991040 Epredia Cytoclip Stainless-Steel Slide Clip Thermo Fisher Scientific Cat#59910052 Glass Coplin Jar Thermo Fisher Scientific Cat#E94 Unifrost Plus Microscope Slides Adhesive, Treated Surfaces Azer Scientific Cat#EMS200G+ ImmEdge Hydrophobic Barrier PAP Pen Vector Laboratories Cat#H-4000 Thermo Scientific Cytospin 4 Cytocentrifuge Thermo Fisher Scientific Cat# A78300003 Sorvall Legend T Benchtop Centrifuge Rotor: 75006445 Sorvall Legend Cat#SO-LEGT BD FACS Aria II sorter BD Biosciences N/A Beckman Coulter Allegra 6 Centrifuge Rotor: GH-3.8 Swing Bucket Rotor Beckman Coulter Cat#BE-A6 Leica TCS SP8 Leica N/A Software and algorithms FlowJo 10 BD http://flowjo.com ll OPEN ACCESSProtocol ..

    FACS:

    Article Title: Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Allophycocyanin/cyanine 7 anti-mouse CD45 (1:100) BioLegend Cat#103116 BV605 anti-mouse CD31 (1:100) BioLegend Cat#102427 (Continued on next page) 2 STAR Protocols 4, 102480, September 15, 2023 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Purified Rat Anti-Mouse CD16/CD32 Clone 2.4G2 (RUO) (1:100) BD Pharmingen Cat #553142 Chemicals, peptides, and recombinant proteins 103 Tris buffered saline (TBS) Bio-Rad Cat#1706463 16% Paraformaldehyde (formaldehyde) aqueous solution, EM grade Thomas Scientific Cat# C993M26 Ammonium chloride (NH4Cl), crystal VWR Chemicals Cat#12125-02-9 Fetal bovine serum (FBS) Gibco Cat#10437-028 DAPI (1:100) BioLegend Cat#422801 PBS (103), pH 7.4 Thermo Fisher Scientific Cat#70011069 PBS (13), pH 7.4 Thermo Fisher Scientific Cat#10010023 Percoll Sigma-Aldrich Cat#17-0891-02 Prolong Gold antifade reagent with DAPI Invitrogen Cat#P36980 RPMI 1640 Medium Thermo Fisher Scientific Cat#11875093 Sodium azide, 5% Fisher Scientific Cat#71448-16 Triton X-100 American Bio Cat#9002-93-1 UltraPure 0.5 M EDTA, pH 8.0 Life Technologies Cat#15575020 Other Cover Glass Thickness 1 1/2 22 3 22 mm Corning Cat#2850-22 Epredia Single Cytofunnel with White Filter Cards Thermo Fisher Scientific Cat#5991040 Epredia Cytoclip Stainless-Steel Slide Clip Thermo Fisher Scientific Cat#59910052 Glass Coplin Jar Thermo Fisher Scientific Cat#E94 Unifrost Plus Microscope Slides Adhesive, Treated Surfaces Azer Scientific Cat#EMS200G+ ImmEdge Hydrophobic Barrier PAP Pen Vector Laboratories Cat#H-4000 Thermo Scientific Cytospin 4 Cytocentrifuge Thermo Fisher Scientific Cat# A78300003 Sorvall Legend T Benchtop Centrifuge Rotor: 75006445 Sorvall Legend Cat#SO-LEGT BD FACS Aria II sorter BD Biosciences N/A Beckman Coulter Allegra 6 Centrifuge Rotor: GH-3.8 Swing Bucket Rotor Beckman Coulter Cat#BE-A6 Leica TCS SP8 Leica N/A Software and algorithms FlowJo 10 BD http://flowjo.com ll OPEN ACCESSProtocol ..

    Software:

    Article Title: Protocol for enrichment, purification, and cytocentrifugation of mouse liver endothelial cells.
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Allophycocyanin/cyanine 7 anti-mouse CD45 (1:100) BioLegend Cat#103116 BV605 anti-mouse CD31 (1:100) BioLegend Cat#102427 (Continued on next page) 2 STAR Protocols 4, 102480, September 15, 2023 Continued REAGENT or RESOURCE SOURCE IDENTIFIER Purified Rat Anti-Mouse CD16/CD32 Clone 2.4G2 (RUO) (1:100) BD Pharmingen Cat #553142 Chemicals, peptides, and recombinant proteins 103 Tris buffered saline (TBS) Bio-Rad Cat#1706463 16% Paraformaldehyde (formaldehyde) aqueous solution, EM grade Thomas Scientific Cat# C993M26 Ammonium chloride (NH4Cl), crystal VWR Chemicals Cat#12125-02-9 Fetal bovine serum (FBS) Gibco Cat#10437-028 DAPI (1:100) BioLegend Cat#422801 PBS (103), pH 7.4 Thermo Fisher Scientific Cat#70011069 PBS (13), pH 7.4 Thermo Fisher Scientific Cat#10010023 Percoll Sigma-Aldrich Cat#17-0891-02 Prolong Gold antifade reagent with DAPI Invitrogen Cat#P36980 RPMI 1640 Medium Thermo Fisher Scientific Cat#11875093 Sodium azide, 5% Fisher Scientific Cat#71448-16 Triton X-100 American Bio Cat#9002-93-1 UltraPure 0.5 M EDTA, pH 8.0 Life Technologies Cat#15575020 Other Cover Glass Thickness 1 1/2 22 3 22 mm Corning Cat#2850-22 Epredia Single Cytofunnel with White Filter Cards Thermo Fisher Scientific Cat#5991040 Epredia Cytoclip Stainless-Steel Slide Clip Thermo Fisher Scientific Cat#59910052 Glass Coplin Jar Thermo Fisher Scientific Cat#E94 Unifrost Plus Microscope Slides Adhesive, Treated Surfaces Azer Scientific Cat#EMS200G+ ImmEdge Hydrophobic Barrier PAP Pen Vector Laboratories Cat#H-4000 Thermo Scientific Cytospin 4 Cytocentrifuge Thermo Fisher Scientific Cat# A78300003 Sorvall Legend T Benchtop Centrifuge Rotor: 75006445 Sorvall Legend Cat#SO-LEGT BD FACS Aria II sorter BD Biosciences N/A Beckman Coulter Allegra 6 Centrifuge Rotor: GH-3.8 Swing Bucket Rotor Beckman Coulter Cat#BE-A6 Leica TCS SP8 Leica N/A Software and algorithms FlowJo 10 BD http://flowjo.com ll OPEN ACCESSProtocol ..



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    Jurkat Lucia Nfat Cd16 Low Cells, supplied by InvivoGen, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 1 article reviews
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    cd16 antibody, anti-human, reafinity  (Miltenyi Biotec)
    93
    Miltenyi Biotec cd16 antibody, anti-human, reafinity
    Characterization and in vitro functional assessment of the MAGE-A4-targeting NKCE. (A) Schematic diagram of the NKCE molecular structure. NKCE is composed of a humanized anti-MAGE-A4 IgG framework fused with an <t>anti-CD16</t> VHH, with an Fc N297A mutation to block Fc mediated ADCC activity. (B) SDS-PAGE analysis under reducing conditions. Lane 1, NKCE; lane 2, 5C4Hu IgG; lane 3, 5C4HuN297A. (C) Thermal stability analysis. The thermal denaturation temperature of the NKCE molecule was determined by circular dichroism (CD). (D) Tumor cell binding affinity. Binding ability of NKCE to HLA-A*02:01 + /MAGE-A4 + tumor cells (NCI-H1703, NCI-H1755, A375, and A375 HLA-A2 overexpress) was measured, CON: blank control with only secondary antibody staining. (E-F) Primary NK cell binding analysis. Flow cytometry demonstrated that NKCE effectively targets the CD3 − CD56 + population in human PBMCs via its anti-CD16 VHH domain. (E) Identification of NK cells in human PBMCs. NK cells were defined as the CD3 − CD56 + population by sequential flow-cytometric gating. (F) NKCE binding was subsequently evaluated within the pre-gated CD3 − CD56 + NK-cell population using a FITC-conjugated anti-human Fc secondary antibody. (G) ELISA comparison of 5C4Hu IgG, 5C4HuN297A, and NKCE binding to the GVY-HLA-A *02:01 complex. (H) Concentration-dependent ELISA binding of NKCE to recombinant CD16a and determination of the apparent EC50. (I) Jurkat-Lucia™ NFAT-CD16a reporter assay evaluating Fc silencing, target-enhanced CD16a reporter activation, and CD16a blockade. The indicated conditions included target cells alone, NKCE without target cells, 5C4Hu IgG with target cells, 5C4HuN297A with target cells, NKCE with target cells, and NKCE with target cells in the presence of a CD16a-blocking antibody. Antibodies were used at 100 nM, and reporter activation was measured after 6 h. Data are presented as mean ± SD of three technical replicate wells from one representative experiment. (J–L) Concentration-dependent cytotoxicity against (J) A375, (K) A375-OVER, and (L) NCI-H1755 cells. Data are presented as mean ± standard error of the mean, n = 3. Statistical significance was assessed using two-way ANOVA followed by multiple-comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.
    Cd16 Antibody, Anti Human, Reafinity, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
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    non ctla 4 expressing jurkat cells  (InvivoGen)
    97
    InvivoGen non ctla 4 expressing jurkat cells
    Characterization and in vitro functional assessment of the MAGE-A4-targeting NKCE. (A) Schematic diagram of the NKCE molecular structure. NKCE is composed of a humanized anti-MAGE-A4 IgG framework fused with an <t>anti-CD16</t> VHH, with an Fc N297A mutation to block Fc mediated ADCC activity. (B) SDS-PAGE analysis under reducing conditions. Lane 1, NKCE; lane 2, 5C4Hu IgG; lane 3, 5C4HuN297A. (C) Thermal stability analysis. The thermal denaturation temperature of the NKCE molecule was determined by circular dichroism (CD). (D) Tumor cell binding affinity. Binding ability of NKCE to HLA-A*02:01 + /MAGE-A4 + tumor cells (NCI-H1703, NCI-H1755, A375, and A375 HLA-A2 overexpress) was measured, CON: blank control with only secondary antibody staining. (E-F) Primary NK cell binding analysis. Flow cytometry demonstrated that NKCE effectively targets the CD3 − CD56 + population in human PBMCs via its anti-CD16 VHH domain. (E) Identification of NK cells in human PBMCs. NK cells were defined as the CD3 − CD56 + population by sequential flow-cytometric gating. (F) NKCE binding was subsequently evaluated within the pre-gated CD3 − CD56 + NK-cell population using a FITC-conjugated anti-human Fc secondary antibody. (G) ELISA comparison of 5C4Hu IgG, 5C4HuN297A, and NKCE binding to the GVY-HLA-A *02:01 complex. (H) Concentration-dependent ELISA binding of NKCE to recombinant CD16a and determination of the apparent EC50. (I) Jurkat-Lucia™ NFAT-CD16a reporter assay evaluating Fc silencing, target-enhanced CD16a reporter activation, and CD16a blockade. The indicated conditions included target cells alone, NKCE without target cells, 5C4Hu IgG with target cells, 5C4HuN297A with target cells, NKCE with target cells, and NKCE with target cells in the presence of a CD16a-blocking antibody. Antibodies were used at 100 nM, and reporter activation was measured after 6 h. Data are presented as mean ± SD of three technical replicate wells from one representative experiment. (J–L) Concentration-dependent cytotoxicity against (J) A375, (K) A375-OVER, and (L) NCI-H1755 cells. Data are presented as mean ± standard error of the mean, n = 3. Statistical significance was assessed using two-way ANOVA followed by multiple-comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.
    Non Ctla 4 Expressing Jurkat Cells, supplied by InvivoGen, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd16/Jurkat-Lucia+NFAT-CD16-Low+Cells/pm42546824-118-0-5
    Average 97 stars, based on 1 article reviews
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    Characterization and in vitro functional assessment of the MAGE-A4-targeting NKCE. (A) Schematic diagram of the NKCE molecular structure. NKCE is composed of a humanized anti-MAGE-A4 IgG framework fused with an anti-CD16 VHH, with an Fc N297A mutation to block Fc mediated ADCC activity. (B) SDS-PAGE analysis under reducing conditions. Lane 1, NKCE; lane 2, 5C4Hu IgG; lane 3, 5C4HuN297A. (C) Thermal stability analysis. The thermal denaturation temperature of the NKCE molecule was determined by circular dichroism (CD). (D) Tumor cell binding affinity. Binding ability of NKCE to HLA-A*02:01 + /MAGE-A4 + tumor cells (NCI-H1703, NCI-H1755, A375, and A375 HLA-A2 overexpress) was measured, CON: blank control with only secondary antibody staining. (E-F) Primary NK cell binding analysis. Flow cytometry demonstrated that NKCE effectively targets the CD3 − CD56 + population in human PBMCs via its anti-CD16 VHH domain. (E) Identification of NK cells in human PBMCs. NK cells were defined as the CD3 − CD56 + population by sequential flow-cytometric gating. (F) NKCE binding was subsequently evaluated within the pre-gated CD3 − CD56 + NK-cell population using a FITC-conjugated anti-human Fc secondary antibody. (G) ELISA comparison of 5C4Hu IgG, 5C4HuN297A, and NKCE binding to the GVY-HLA-A *02:01 complex. (H) Concentration-dependent ELISA binding of NKCE to recombinant CD16a and determination of the apparent EC50. (I) Jurkat-Lucia™ NFAT-CD16a reporter assay evaluating Fc silencing, target-enhanced CD16a reporter activation, and CD16a blockade. The indicated conditions included target cells alone, NKCE without target cells, 5C4Hu IgG with target cells, 5C4HuN297A with target cells, NKCE with target cells, and NKCE with target cells in the presence of a CD16a-blocking antibody. Antibodies were used at 100 nM, and reporter activation was measured after 6 h. Data are presented as mean ± SD of three technical replicate wells from one representative experiment. (J–L) Concentration-dependent cytotoxicity against (J) A375, (K) A375-OVER, and (L) NCI-H1755 cells. Data are presented as mean ± standard error of the mean, n = 3. Statistical significance was assessed using two-way ANOVA followed by multiple-comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

    Journal: mAbs

    Article Title: Artificial intelligence-assisted design of MAGE-A4 ×CD16 T-cell receptor-like natural killer cell engagers

    doi: 10.1080/19420862.2026.2717991

    Figure Lengend Snippet: Characterization and in vitro functional assessment of the MAGE-A4-targeting NKCE. (A) Schematic diagram of the NKCE molecular structure. NKCE is composed of a humanized anti-MAGE-A4 IgG framework fused with an anti-CD16 VHH, with an Fc N297A mutation to block Fc mediated ADCC activity. (B) SDS-PAGE analysis under reducing conditions. Lane 1, NKCE; lane 2, 5C4Hu IgG; lane 3, 5C4HuN297A. (C) Thermal stability analysis. The thermal denaturation temperature of the NKCE molecule was determined by circular dichroism (CD). (D) Tumor cell binding affinity. Binding ability of NKCE to HLA-A*02:01 + /MAGE-A4 + tumor cells (NCI-H1703, NCI-H1755, A375, and A375 HLA-A2 overexpress) was measured, CON: blank control with only secondary antibody staining. (E-F) Primary NK cell binding analysis. Flow cytometry demonstrated that NKCE effectively targets the CD3 − CD56 + population in human PBMCs via its anti-CD16 VHH domain. (E) Identification of NK cells in human PBMCs. NK cells were defined as the CD3 − CD56 + population by sequential flow-cytometric gating. (F) NKCE binding was subsequently evaluated within the pre-gated CD3 − CD56 + NK-cell population using a FITC-conjugated anti-human Fc secondary antibody. (G) ELISA comparison of 5C4Hu IgG, 5C4HuN297A, and NKCE binding to the GVY-HLA-A *02:01 complex. (H) Concentration-dependent ELISA binding of NKCE to recombinant CD16a and determination of the apparent EC50. (I) Jurkat-Lucia™ NFAT-CD16a reporter assay evaluating Fc silencing, target-enhanced CD16a reporter activation, and CD16a blockade. The indicated conditions included target cells alone, NKCE without target cells, 5C4Hu IgG with target cells, 5C4HuN297A with target cells, NKCE with target cells, and NKCE with target cells in the presence of a CD16a-blocking antibody. Antibodies were used at 100 nM, and reporter activation was measured after 6 h. Data are presented as mean ± SD of three technical replicate wells from one representative experiment. (J–L) Concentration-dependent cytotoxicity against (J) A375, (K) A375-OVER, and (L) NCI-H1755 cells. Data are presented as mean ± standard error of the mean, n = 3. Statistical significance was assessed using two-way ANOVA followed by multiple-comparison test. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

    Article Snippet: To evaluate the silencing of ADCC activity by the N297A mutation, an ADCC reporter assay was performed using Jurkat-LuciaTM NFAT-CD16 effector cells (InvivoGen, Cat. Code: jktl-nfat-cd16).

    Techniques: In Vitro, Functional Assay, Mutagenesis, Blocking Assay, Activity Assay, SDS Page, Circular Dichroism, Binding Assay, Control, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Comparison, Concentration Assay, Recombinant, Reporter Assay, Activation Assay