Cd19 fitc clone 1d3

Manufactured by BD

Sourced in Germany

The CD19 FITC (clone 1D3) is a fluorochrome-conjugated monoclonal antibody that binds to the CD19 antigen expressed on the surface of B cells. It is a tool used in flow cytometry applications for the identification and enumeration of B cells.

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Lab products found in correlation

Cd3e percp efluor 710 clone 17a2, by Thermo Fisher Scientific (4 mentions) Cd11c apc hl3, by BD (4 mentions) Cd11b pe cy7 clone m1 70, by Thermo Fisher Scientific (4 mentions) Facsaria 3, by BD (4 mentions) Hla dr bv605 clone g46 6, by BD (3 mentions) Fc block, by BD (3 mentions) B220 apc clone ra3 6b2, by BD (1 mentions) Cd11c apc clone hl3, by BD (1 mentions) Pd 1 pe clone rmp1 30, by BioLegend (1 mentions) Lsr 2, by BD (1 mentions) Ethylenediaminetetraacetic acid, by Thermo Fisher Scientific (1 mentions) Potassium bicarbonate, by Merck Group (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Hla dr bv605, by BD (1 mentions) 70 μm nylon cell strainer, by BD (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) β mercaptoethanol, by Merck Group (1 mentions)

Spelling variants of this product

CD19-FITC (75 citations) CD19 (1D3, (23 citations) CD19 (clone 1D3, (15 citations) Clone 1D3 (5 citations) CD19 FITC (1D3), (4 citations) FITC-conjugatedanti-mouse CD19 (clone 1D3) (2 citations)

5 protocols using cd19 fitc clone 1d3

Splenic B Cell Isolation Protocol

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Splenocytes were prepared as described before. Afterwards, cells were stained for CD3e PerCP-eFluor 710 (clone 17A2, Thermo Fisher Scientific), CD19 FITC (clone 1D3, BD Biosciences), CD11c APC (HL3, BD Biosciences), CD11b PE-Cy7 (clone M1/70, Thermo Fisher Scientific) and HLA-DR BV605 (clone G46-6, BD Biosciences) and subsequently sorted using a BD FACS Aria III (BD Biosciences). Post sort analysis using FlowJo software (Version 10.8.1; BD Biosciences) showed a purity of >98% for splenic CD19+ B cells.

Lubich C., Steinitz K.N., Hoelbl B., Prenninger T., van Helden P.M., Weiller M, & Reipert B.M. (2022). Modulating the microenvironment during FVIII uptake influences the nature of FVIII-peptides presented by antigen-presenting cells. Frontiers in Immunology, 13, 975680.

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Flow Cytometric Characterization of Immune Cells

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Cells were stained for flow cytometry at 4° C for 30 minutes in FACS buffer (PBS, 1% BSA, and 1mM EDTA) plus appropriate antibodies and fixed in 0.1% paraformaldehyde for at least 1 hour. Magnetically sorted B cells were gated on CD19+ B220+ CD11b− and CD11c− events. Magnetically sorted CD4 T cells were gated on CD3+ CD8− CD4+ CD11b− and CD11c− events. Primary BMDMs were gated on CD11b+ CD11c− MHCII+ cells. RAW 264.7 macrophages, primary BMDMs, T cells, and B cells were also stained with anti-PD-1 antibody for analysis. Antibodies used were CD4 PerCP-Cy5.5 (clone RM45, Tonbo Biosciences Corp., San Diego, CA), CD8 FITC (clone 53-6.7, Tonbo Biosciences Corp., San Diego, CA), CD19 FITC (clone 1D3, BD Pharmingen, San Diego, CA), B220 APC (clone RA3-6B2, BD Pharmingen, San Diego, CA), CD11b FITC (clone M1/70 BD Pharmingen, San Diego, CA), CD11C APC (clone HL3, BD Pharmingen, San Diego, CA), PD-1 PE (clone RMP1-30, Biolegend Inc., San Diego, CA). Flow cytometry was performed on a BD LSR-II and analyzed using FlowJo 9.6.4 software.

Bally A.P., Lu P., Tang Y., Austin J.W., Scharer C.D., Ahmed R, & Boss J.M. (2015). NF-κB regulates PD-1 expression in macrophages. Journal of immunology (Baltimore, Md. : 1950), 194(9), 4545-4554.

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Isolation of Splenic Dendritic Cells

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Splenocytes were prepared as described before. Dendritic cells were enriched by a pre-isolation step using NKp46, CD3e and CD19 MACS beads negative selection (all Miltenyi Biotech, Germany). Subsequently, cells were stained for CD3e PerCP-eFluor 710 (clone 17A2, Thermo Fisher Scientific), CD19 FITC (clone 1D3, BD Biosciences), CD11c APC (HL3, BD Biosciences), CD11b PE-Cy7 (clone M1/70, Thermo Fisher Scientific) and HLA-DR BV605 (clone G46-6, BD Biosciences) and sorted using a BD FACS Aria III (BD Biosciences). Nonspecific binding through Fc gamma receptors was blocked by a mixture of anti-CD16 and anti-CD32 antibodies (Fc Block; BD Biosciences). Post sort analysis using FlowJo software (Version 10.8.1; BD Biosciences) showed an average purity of 94.8% for splenic CD11c high MHC class II+ dendritic cells.

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Profiling Antigen-Presenting Cells in Murine Spleen

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Spleens were finely minced and passed through a 70-μM nylon cell strainer (Becton Dickinson, Franklin Lakes, NJ). Spleen cells were collected in RPMI 1640 (Life Technologies, Paisley, Scotland) supplemented with 10% preselected fetal calf serum (FCS), 2 mM l-glutamine (both from Hyclone, Logan, UT), 100 U/mL penicillin, 100 mg/mL streptomycin (both from Life Technologies), and 5 × 10–5 M β-mercaptoethanol (Sigma-Aldrich). Single-cell suspensions were cleared of erythrocytes by hemolysis using a hypotonic buffer (pH 7.2) composed of 0.15 M ammonium chloride, 10 mM potassium bicarbonate (both from Merck, Darmstadt, Germany), and 0.1 mM ethylene-diaminetetraacetic acid (Life Technologies).
To evaluate the composition of APCs, splenocytes were stained for CD3e PerCP-eFluor 710 (clone 17A2, Thermo Fisher Scientific), CD19 FITC (clone 1D3, BD Biosciences), CD11c APC (HL3, BD Biosciences), CD11b PE-Cy7 (clone M1/70, Thermo Fisher Scientific) and HLA-DR BV605 (clone G46-6, BD Biosciences) and analyzed using a BD FACS Aria III (BD Biosciences) and FlowJo software (Version 10.8.1; BD Biosciences). Nonspecific binding through Fc gamma receptors was blocked by a mixture of anti-CD16 and anti-CD32 antibodies (Fc Block; BD Biosciences).

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Enrichment and Characterization of Splenic Monocytes

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Splenocytes were prepared as described before. Monocytes/macrophages were enriched by a pre-isolation step using NKp46, CD3e and CD19 MACS beads negative selection (all Miltenyi Biotech, Germany). Subsequently, cells were stained for CD3e PerCP-eFluor 710 (clone 17A2, Thermo Fisher Scientific), CD19 FITC (clone 1D3, BD Biosciences), CD11c APC (HL3, BD Biosciences), CD11b PE-Cy7 (clone M1/70, Thermo Fisher Scientific) and HLA-DR BV605 (clone G46-6, BD Biosciences) and sorted using a BD FACS Aria III (BD Biosciences). Nonspecific binding through Fc gamma receptors was blocked by a mixture of anti-CD16 and anti-CD32 antibodies (Fc Block; BD Biosciences). Post sort analysis using FlowJo software (Version 10.8.1; BD Biosciences) showed an average purity of 94.4% for splenic CD11b+ monocytes/macrophages.

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