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Cd19 clone 1d3

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The CD19 (clone 1D3) is a laboratory reagent used for the identification and enumeration of CD19-positive cells, such as B lymphocytes, in biological samples. It is a monoclonal antibody that specifically binds to the CD19 surface antigen expressed on B cells. This reagent can be used in flow cytometry and other immunoassay techniques to quantify the presence and proportion of CD19-positive cells in a given sample.

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Lab products found in correlation

Anti mouse cd16 cd32, by BD (2 mentions) Facscanto 2 cytometer, by BD (2 mentions) Live dead fixable dead cell stain, by Thermo Fisher Scientific (2 mentions) B220 clone ra3 6b2, by Thermo Fisher Scientific (2 mentions) Cd11b clone m1 70, by BioLegend (2 mentions) C kit clone 2b8, by Thermo Fisher Scientific (1 mentions) Cd43 clone s7, by BD (1 mentions) Igd clone ia6 2, by BioLegend (1 mentions) Clone rb6 8c5, by Thermo Fisher Scientific (1 mentions) Cd3 clone 145 2c11, by Thermo Fisher Scientific (1 mentions) Mac3 m3 84, by BD (1 mentions) Zombie uv fixable viability dye, by BioLegend (1 mentions) Cd45.2 clone 104, by BioLegend (1 mentions) Gr 1 clone rb6 8c5, by BioLegend (1 mentions) Lymphoprep, by STEMCELL (1 mentions) Gentlemacs octo dissociator, by Miltenyi Biotec (1 mentions) Cd4 clone rm4 5, by BioLegend (1 mentions) Nk1.1 clone pk136, by BioLegend (1 mentions) Collagenase, by Merck Group (1 mentions) Cd45 clone 104, by BioLegend (1 mentions)

Close spelling variants of this product

CD19 (1D3, (22 citations) Anti-CD19 (clone 1D3), (2 citations) Anti-CD19–PE-Cy7 (clone eBio1D3 (1D3), (2 citations) CD19 PE (clone 1D3, (2 citations) Anti-mouse CD19(clone 1D3, (2 citations)

8 protocols using cd19 clone 1d3

1

Immune Cell Profiling by Flow Cytometry

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Cells were stained and analyzed by flow cytometry as previously described (Baxter and Griffin, 2016 (link)). Briefly, 106 live cells were stained with violet LIVE/DEAD Fixable Dead Cell Stain (Life Technologies) for 30 min, anti-mouse CD16/CD32 (BD Pharmingen) for 15 min, and monoclonal antibodies against CD45 (clone 30-F11), CD3 (clone 17A2), CD4 (clone RM4-5), CD8a (clone 53-6.7), and CD19 (clone 1D3) from Ebioscience or BD Pharmingen for 30 min. Cells were run on a BD FACSCanto II cytometer using BD FACSDiva software, version 8, and analyses were carried out using FlowJo software, version 8. Cells were characterized as follows: CD4 T cells (CD45hiCD3+CD4+), CD8 T cells (CD45hiCD3+CD8+), and B cells (CD45hiCD3-CD19+). For CLN and brain, total mononuclear cell counts were from three independent experiments, and for spinal cord, as well as CD4+, CD8+ and CD19+ cells for all tissues, counts were from two independent experiments. At 11 DPI, the SINV, 0.6 mg/kg DON group had data from one fewer experiment than the others (one to two independent experiments).
Baxter V.K., Glowinski R., Braxton A.M., Potter M.C., Slusher B.S, & Griffin D.E. (2017). Glutamine antagonist-mediated immune suppression decreases pathology but delays virus clearance in mice during nonfatal alphavirus encephalomyelitis. Virology, 508, 134-149.
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2

Flow Cytometric Analysis of Murine Bone Marrow

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Eight-week-old mice were used for FACS analyses of the BM. Single BM cell suspensions were stained using anti-mouse Abs against B220 (clone RA3-6B2; eBioscience), CD19 (clone 1D3; eBioscience), CD43 (clone S7; BD Biosciences, Franklin Lakes, NJ), IgM (clone RMM-1; BioLegend), IgD (clone IA6-2; BioLegend), c-Kit (clone 2B8; eBioscience), Flk2 (clone A2F10; eBioscience) and Ter119 (clone Ter119; eBioscience). The gating was done as described (17 (link), 18 ).
Catalan-Dibene J., Vazquez M.I., Luu V.P., Nuccio S.P., Karimzadeh A., Kastenschmidt J.M., Villalta S.A., Ushach I., Pone E.J., Casali P., Raffatellu M., Burkhardt A.M., Hernandez-Ruiz M., Heller G., Hevezi P.A, & Zlotnik A. (2017). Identification of IL-40, a Novel B Cell–Associated Cytokine. Journal of immunology (Baltimore, Md. : 1950), 199(9), 3326-3335.
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3

Quantifying Airway Inflammatory Cells by Flow Cytometry

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IL-4, IL-5, IL-13, IgE and IgG1 in BALF were measured using commercially available enzyme-linked immunosorbent assay (ELISA) kits from eBioscience according to the manufacturer’s recommendations. Flow cytometry was used to test the percentages of the various inflammatory cells in the BAL. Eosinophils were defined as SSChigh SiglecF+ (clone E50-2440, BD) Mac-3− (M3/84, BD) cells, alveolar macrophages were identified as SSChigh SiglecF+ Mac-3+ cells, granulocytes were recognized as SSChigh Gr-1+ (clone RB6-8C5, eBioscience) cells, and lymphocytes were identified as FSClow/SSClow and expressing CD3 (clone 145-2C11, eBioscience) or CD19 (clone 1D3, eBioscience).
Niu R., Xiao X., Liu B., Li Y., zhong Y, & Ma L. (2017). RETRACTED ARTICLE: Inhibition of airway inflammation in a cockroach allergen model of asthma by agonists of miRNA-33b. Scientific Reports, 7, 7409.
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4

Flow Cytometry Analysis of Immune Cells

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Cells were stained and analyzed by flow cytometry as previously described (Baxter and Griffin, 2016 (link)). Briefly, 106 live cells were stained with violet LIVE/DEAD Fixable Dead Cell Stain (Life Technologies) for 30 min, anti-mouse CD16/CD32 (BD Pharmingen) for 15 min, and monoclonal antibodies against CD45 (clone 30-F11), CD3 (clone 17A2), CD4 (clone RM4-5), CD8a (clone 53-6.7), and CD19 (clone 1D3) from Ebioscience or BD Pharmingen for 30 min. Cells were run on a BD FACSCanto II cytometer using BD FACSDiva software, version 8, and analyses were carried out using FlowJo software, version 8. Cells were characterized as follows: CD4 T cells (CD45hiCD3+CD4+), CD8 T cells (CD45hiCD3+CD8+), and B cells (CD45hiCD3-CD19+). For CLN and brain, total mononuclear cell counts were from three independent experiments and for spinal cord, as well as CD4+, CD8+ and CD19+ cells for all tissues, counts were from two independent experiments. At 11 DPI, the SINV, 0.6mg/kg DON group had data from one fewer experiment than the others (one to two independent experiments).
Baxter V.K., Glowinski R., Braxton A.M., Potter M.C., Slusher B.S, & Griffin D.E. (2017). Glutamine antagonist-mediated immune suppression decreases pathology but delays virus clearance in mice during nonfatal alphavirus encephalomyelitis. Virology, 508, 134-149.
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5

Tumor Dissociation and Immune Cell Sorting

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Tumors were harvested on day 5 and homogenized in RPMI using a gentleMACS Octo Dissociator (Miltenyi Biotec) on m_lung_01_01 and m_lung_02_01settings. Cells were separated with Lymphoprep (Stem Cell Technologies) according to the manufacturer’s instructions and stained for sorting with Zombie UV Fixable Viability Dye (BioLegend) as well as antibodies specific for CD45.2 (clone 104; Biolegend), CD11b (clone M1/70; Biolegend), and Gr-1 (clone RB6-8C5; Biolegend), CD4 (clone RM4-5; Biolegend), CD19 (clone 1D3; eBiosciences), and NK1.1 (clone PK136; Biolegend). Representative FACS plots from sorting are shown in Supplemental Figure 4.
Wilski N.A., Stotesbury C., Del Casale C., Montoya B., Wong E., Sigal L.J, & Snyder C.M. (2020). STING sensing of Murine Cytomegalovirus alters the tumor microenvironment to promote anti-tumor immunity. Journal of immunology (Baltimore, Md. : 1950), 204(11), 2961-2972.
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6

Detailed Splenocyte Isolation and Characterization

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Splenocytes were isolated by digesting sliced spleens in 0.1 U/mL collagenase (Sigma Aldrich) in Hank’s Balanced Salt Solution (Corning) for 30 minutes at 37°C. Digestion was stopped by addition of EDTA to 5 mM, and the resulting mixture was passed through a cell strainer. Red blood cells were removed with ACK lysing buffer (Lonza), and cells were washed in FACS buffer (PBS supplemented with 2% FBS and 2 mM EDTA; Corning and HyClone) and re-strained.
Cells were stained by conventional methods in FACS buffer as previously described43 (link). Antibodies used for staining were CD19 (clone 1D3, eBioscience), CD3 (clone 17A2, eBioscience), CD31 (clone 390, BioLegend), CD45 (clone 104, BioLegend), NK1.1 (clone PK136, eBioscience), CD11b (clone M1/70, BioLegend), CD11c (clone N418, BioLegend), Siglec H (clone 551, BioLegend), F4/80 (clone BM8, BioLegend), and Ly-6G (clone 1A8, BioLegend). Cells were also stained with LIVE/DEAD viability dyes (Thermo Fisher) to exclude dead cells.
Described splenocyte cell types (Fig. 5A, Fig. S5C) from four mice were isolated by FACS on two FACSAria II cell sorters (BD Biosciences) at the Emory University School of Medicine Flow Cytometry Core.
Paunovska K., Sago C.D., Monaco C.M., Hudson W.H., Castro M.G., Rudoltz T.G., Kalathoor S., Vanover D.A., Santangelo P.J., Ahmed R., Bryksin A.V, & Dahlman J.E. (2018). A direct comparison of in vitro and in vivo nucleic acid delivery mediated by hundreds of nanoparticles reveals a weak correlation. Nano letters, 18(3), 2148-2157.
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7

Multiparameter flow cytometric analysis of immune cells

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Single cell suspensions were incubated with 100 µL diluted antibodies for 30 min at 4°C in ice-cold fluorescence-activated cell sorting (FACS) buffer (0.5% bovine serum albumin, 2 mM EDTA in PBS) with ‘cocktails’ of the following antibodies: CD45 clone 30-F11, CD3e clone 145–2 C11, CD4 clone RM4-5, CD62L clone MEL-14, CD44 clone IM7, CD8a clone 53–6.7, B220 clone RA3-6B2, CD23 clone B3B4, CD19 clone 1D3 and CD5 clone 53–7.3 (all from eBiosciences), CD21 clone 7G6 (BD biosciences) and CD11c clone N418, F4/80 clone BM8, CD64 clone X54-5/7.1 and NK1.1 clone PK136. Intracellular staining for Ki67 clone B56 (BD Biosciences) and pS6 PE and pAKT Alexa Fluor 488 (Cell signalling) was performed by using the Cytofix/Perm kit (BD biosciences) and Fixation/Permeabilization Buffer set (ebiosciences) according to the manufacturer’s protocol. Cells were resuspended in FACS buffer and then analysed using a Cyan-ADP (Dako) or Fortessa (BD) with forward/side scatter gates set to exclude non-viable cells. Cells of interest were sorted by using BD FACSAria. Data were analysed with FlowJo software (Tree Star).
Nayar S., Campos J., Smith C.G., Iannizzotto V., Gardner D.H., Colafrancesco S., Pipi E., Kollert F., Hunter K.J., Brewer C., Buckley C.D., Bowman S.J., Priori R., Valesini G., Juarez M., Fahy W.A., Fisher B.A., Payne A., Allen R.A, & Barone F. (2018). Phosphatidylinositol 3-kinase delta pathway: a novel therapeutic target for Sjögren’s syndrome. Annals of the Rheumatic Diseases, 78(2), 249-260.
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8

Characterization of Peritoneal Leukocytes

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Mice untreated or treated with intraperitoneal injection of 1 mL 4% thioglycolate (Sigma-Aldrich) for 4, 24, or 48 h were killed using CO 2 , and peritoneal leukocytes were collected by lavage of the cavity with PBS containing 5 mM EDTA. After centrifugation, cells were incubated 2 min in 1 mL of RBC buffer (170 mM NH4Cl, 12 mM KHCO3, 0.9 mM EDTA, pH 7.6) to lyse residual red cells and, then, centrifugued and resuspended in DMEM and counted using a hemocytometer. For flow cytometry analysis, PerC cells were incubated for 30 min at 4ºC with labeled antibodies against CD19 (clone 1D3, eBioscience), F4/80 (clone Cl:A3-1, AbD Serotec), Ly6-G (clone RB6-8C5, eBioscience), CD5 (clone 53-7.3, Biolegend), CD11b (clone M1/70, BD Pharmingen), and CD3ε (clon 145-2C11, BD Pharmingen), using isotype-matched antibodies as negative control and color compensation as appropriate for dual labeling. Samples were analyzed in a Coulter flow cytometer model FC500. A more detailed analysis of B-cell population was performed by simultaneous labeling with anti-CD19, anti-CD5 and anti-CD11b antibodies. According to the expression of these markers, B cells were considered as belonging to B1a (CD19+/CD5+/CD11b+), B1b (CD19+/CD5-/CD11b+), B1c (CD19+/CD5+/CD11b-) or B2 (CD19+/CD5-/CD11b-) subsets (Tung and Herzenberg, 2007; Rosado et al., 2014; Hastings et al., 2006) .
Porras G., Ayuso M.S, & González-Manchón C. (2018). Leukocyte-endothelial cell interaction is enhanced in podocalyxin-deficient mice. The international journal of biochemistry & cell biology, 99.
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