rat mab anti cd90  (R&D Systems)

Journal: Journal of Virology

Article Title: Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry

doi: 10.1128/JVI.01742-17

Figure Lengend Snippet: Inhibition of HCVcc entry into Huh7.5 cells by anti-HAVCR1 antibody. Huh7.5 cells were treated with decreasing concentrations of the following antibodies following 2-fold dilutions from 4 μg/ml to 0.0625 μg/ml: anti-HAVCR1 MAb (1D12) (A), anti-CD81 MAb (JS-81) (B), and isotype control (C). Cells were treated with antibodies 1 h prior to infection with J6/JFH1. After 72 h, HCVcc infection was evaluated by automatic counting of stained foci and the percentage of inhibition calculated by comparing focus counts in treated and mock-treated cells. (D) Huh7.5 cells were treated with increasing concentrations of anti-CD81 and anti-HAVCR1 MAbs individually or in combination. The percentage of inhibition and dose effect plot was created using CalcuSyn 2.0 software. Testing for all assays was performed in duplicate. Error bars represent standard errors of the means. Data are representative of three independent experiments.

Article Snippet: To characterize receptor expression on the surface of Huh7.5 cells, we performed staining with the following antibodies: anti-CD81 mouse MAb (JS-81; BD Pharmingen, San Jose, CA); anti-SRB1 antibody (m1B9; BioLegend, San Diego, CA), anti-HAVCR1 phycoerythrin (PE)-labeled mouse MAb (REA384; Miltenyi Biotec, Inc., Auburn, CA), or purified HAVCR1-1 MAb, an antibody directed against the IgV domain of HAVCR1 that blocks binding of phospholipids.

Techniques: Inhibition, Control, Infection, Staining, Software

Journal: Journal of Virology

Article Title: Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry

doi: 10.1128/JVI.01742-17

Figure Lengend Snippet: Comparison of inhibitory activities and cell membrane localization of CD81 and HAVCR1. JFH1-Nanoluc was added to cells at 4°C for 1 h before moving the cells to 37°C to allow entry and infection to proceed. Antibody against CD81 (JS-81) (A), HAVCR1 (1D12) (B), or mouse isotype control (C) was added hourly to cells before and after HCVcc binding. The amount of antibody used was the concentration that was previously found to result in 50% inhibition of infection or the highest equivalent concentration in the case of the isotype control. The levels of luciferase expression at different time points were measured, and the percentages of inhibition were calculated by comparing the signals in mock-treated and infected cells at each time point. Error bars represent standard errors of the means. (D) Confocal analysis of Huh7.5 cells surface stained with 4′,6-diamidino-2-phenylindole (DAPI), anti-CD81 MAb (JS-81), and anti-HAVCR1 MAb (HAVCR1-1 MAb).

Article Snippet: To characterize receptor expression on the surface of Huh7.5 cells, we performed staining with the following antibodies: anti-CD81 mouse MAb (JS-81; BD Pharmingen, San Jose, CA); anti-SRB1 antibody (m1B9; BioLegend, San Diego, CA), anti-HAVCR1 phycoerythrin (PE)-labeled mouse MAb (REA384; Miltenyi Biotec, Inc., Auburn, CA), or purified HAVCR1-1 MAb, an antibody directed against the IgV domain of HAVCR1 that blocks binding of phospholipids.

Techniques: Comparison, Membrane, Infection, Control, Binding Assay, Concentration Assay, Inhibition, Luciferase, Expressing, Staining

Journal: Journal of Virology

Article Title: Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry

doi: 10.1128/JVI.01742-17

Figure Lengend Snippet: Dose-dependent inhibition of HCVcc entry with soluble HAVCR1 receptors, phosphatidyl serine, or annexin V. HCVcc was incubated 1 h prior to infection of Huh7.5 cells with increasing amounts of soluble HAVCR1 receptor containing the IgV- and mucinlike domains (HAVCR1-Fc) (A), soluble monkey HAVCR1 (mkHAVCR1-Fc) or mouse Havcr1 ortholog (mHavcr1-Fc) (B), HAVCR1-Fc or soluble HAVCR1 containing the IgV-like domain [HAVCR1(IgV)-Fc] (C), HAVCR1-Fc or the same receptor with an N94A mutation in the phosphatidylserine binding domain (HAVCR1-N94A-Fc) (D), phosphatidylserine (PS) or phosphatidylcholine (PC) liposomes (E), or annexin V (F). Testing for all assays was performed in duplicate. Error bars represent standard errors of the means. Data are representative of three independent experiments. All experiments were performed with HCVcc J6/JFH1 except for that shown in panel A, which used HCVcc chimeric viruses 1a/2a, 1b/2a, 2a/2a (J6/JFH1), and 3a/2a.

Article Snippet: To characterize receptor expression on the surface of Huh7.5 cells, we performed staining with the following antibodies: anti-CD81 mouse MAb (JS-81; BD Pharmingen, San Jose, CA); anti-SRB1 antibody (m1B9; BioLegend, San Diego, CA), anti-HAVCR1 phycoerythrin (PE)-labeled mouse MAb (REA384; Miltenyi Biotec, Inc., Auburn, CA), or purified HAVCR1-1 MAb, an antibody directed against the IgV domain of HAVCR1 that blocks binding of phospholipids.

Techniques: Inhibition, Incubation, Infection, Mutagenesis, Binding Assay, Liposomes

Journal: Journal of Virology

Article Title: Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry

doi: 10.1128/JVI.01742-17

Figure Lengend Snippet: Reduced HAVCR1 expression on Huh7.5 cells results in reduced levels of HCVcc and HCVpp entry. Huh7.5 HAVCR1-1 knockdown (KD) cells were generated by transfection of parent Huh7.5 cells with a plasmid expressing HAVCR1 shRNA. (A) Percentages of parent and KD cells positive for HAVCR1, CD81, and SRB1 surface expression. Bars represent the mean values from 5 independent experiments for HAVCR1 and CD81 and 4 independent experiments for SRB1. (B) Mean fluorescence signals of HAVCR1, CD81, and SRB1 surface expression on parent and KD cells. Bars represent the mean values from 5 independent experiments for HAVCR1 and CD81 and 4 independent experiments for SRB1. (C) Western blot detection of claudin and occludin after cell surface biotinylation of Huh7.5 and HAVCR1 KD cells. Whole-cell lysate (WCL) is cell lysate prior to pulldown treatment. Nonbiotinylated control represents cells treated exactly as for the biotinylated cells but without the addition of EZ-Link sulfo-NHS-LCLC-biotin for cell surface biotinylation. Anti-claudin antibody was used for detection and shows the specificity of the NeutrAvidin beads for precipitation. (D) Luciferase activities at 24, 48, and 72 h postinfection in lysates of parent and KD cells infected with JFH1-Nanoluc. Bars represent the mean relative light units (RLU) calculated from 4 independent experiments using 9 replicates per experiment. (E) Titers of virus recovered from the supernatants of parent and KD cells at 72 h postinfection with JFH1-Nanoluc. Bars represent the mean titers from 4 independent experiments. (F) Luciferase activities in parent and KD cells 48 h after infection with HCVpp. (G) Luciferase activities in parent and KD cells 48 h after infection with VSV-G. Bars represent the mean values from 4 independent experiments. Error bars in all graphs represent standard errors of the means. Statistical analyses were performed using the Mann-Whitney test.

Article Snippet: To characterize receptor expression on the surface of Huh7.5 cells, we performed staining with the following antibodies: anti-CD81 mouse MAb (JS-81; BD Pharmingen, San Jose, CA); anti-SRB1 antibody (m1B9; BioLegend, San Diego, CA), anti-HAVCR1 phycoerythrin (PE)-labeled mouse MAb (REA384; Miltenyi Biotec, Inc., Auburn, CA), or purified HAVCR1-1 MAb, an antibody directed against the IgV domain of HAVCR1 that blocks binding of phospholipids.

Techniques: Expressing, Knockdown, Generated, Transfection, Plasmid Preparation, shRNA, Fluorescence, Western Blot, Control, Luciferase, Infection, Virus, MANN-WHITNEY

Journal: Journal of Virology

Article Title: Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry

doi: 10.1128/JVI.01742-17

Figure Lengend Snippet: Complete silencing of HAVCR1 expression in Huh7.5 cells significantly reduces HCVcc infection. Surface expression levels of receptors on Huh7.5 cells, Huh7-A-I cells, and different clones of Huh7-A-I HAVCR1 knockout (KO) cells were assessed by FACS using MAbs specific to HAVCR1 (REA383) (A), CD81 (JS-81) (B), and SRB1 (m1B9) (C). Data for positive cells are shown in dark gray, and data for mouse PE-isotype control are shown in light gray. Data are representative of three independent experiments. (D) Western blot detection of claudin and occludin after cell surface biotinylation of Huh7-A-I and clone 12 KO cells. Whole-cell lysate (WCL) is cell lysate prior to pulldown treatment. Nonbiotinylated control represents cells treated exactly as for the biotinylated cells but without the addition of EZ-Link sulfo-NHS-LCLC-biotin for cell surface biotinylation. Anticlaudin antibody was used for detection and shows the specificity of the NeutrAvidin beads for precipitation. Antiactin antibody was used for detection of actin, a cytoplasmic protein that is not expressed on the cell surface, demonstrating biotinylation of surface proteins only. (E) Luciferase activities in lysates from Huh7-A-I HAVCR1 KO clones and parent cell line at 72 h postinfection with JFH1-Nanoluc. Data were obtained from 3 independent experiments, and each experiment was carried out using 12 replicates for each cell clone. Each symbol represents an individual well from the 3 independent experiments. Horizontal bars represent the mean luciferase activities. P values were calculated using the mean values from each of the 3 independent experiments. (F) Time course of luciferase expression relative to the expression at 3 h posttransfection in Huh7-A-I HAVCR1 KO clones 2 and 12 and parent cells following transfection with JFH1-Nanoluc RNA transcribed in vitro. Data are the mean values from 6 replicates at each time point. Error bars represent standard errors of the means. Data are representative of two independent experiments. RLU, relative light units.

Article Snippet: To characterize receptor expression on the surface of Huh7.5 cells, we performed staining with the following antibodies: anti-CD81 mouse MAb (JS-81; BD Pharmingen, San Jose, CA); anti-SRB1 antibody (m1B9; BioLegend, San Diego, CA), anti-HAVCR1 phycoerythrin (PE)-labeled mouse MAb (REA384; Miltenyi Biotec, Inc., Auburn, CA), or purified HAVCR1-1 MAb, an antibody directed against the IgV domain of HAVCR1 that blocks binding of phospholipids.

Techniques: Expressing, Infection, Clone Assay, Knock-Out, Control, Western Blot, Luciferase, Transfection, In Vitro

Journal: Journal of Virology

Article Title: Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry

doi: 10.1128/JVI.01742-17

Figure Lengend Snippet: Expression of HAVCR1 in KO clone 12 enhances HCV infection. HAVCR1 KO clone 12 cells were transfected with plasmids expressing HAVCR1, mutated forms of HAVCR1 (E88A, N94A, Y330A, Y336A, and del324–331 mutations), or vector. (A) Expression of HAVCR1 on the surface of clone 12 cells transfected with vector or plasmid expressing HAVCR1 48 h posttransfection was assessed using anti-HAVCR1-1 MAb. (B) Apoptotic binding activities of clone 12 cells transfected with vector or plasmid expressing HAVCR1. (C) Luciferase expression in cell lysates of clone 12 cells transfected with HAVCR1 expression plasmid and infected with JFH1-Nanoluc. Data at 24 h postinfection are shown. Four independent experiments were performed; each time point included 3 replicates. (D) Expression of E88A or N94A mutant on the surface of transfected clone 12 cells 48 h posttransfection assessed using anti-HAVCR1-1 MAb. (E) Apoptotic binding of clone 12 cells transfected with vector or plasmids expressing E88A and N94A mutant forms of HAVCR1. (F) Luciferase expression in cell lysates of clone 12 cells transfected with plasmids expressing E88A or N94A mutants and infected with JFH1-Nanoluc. Data are from 4 independent experiments, each including 3 replicates, at 24 h postinfection. (G) Expression of Y330A, Y336A, or del324–331 mutant on the surface of transfected clone 12 cells 48 h posttransfection was assessed using anti-HAVCR1-1 MAb. (H) Apoptotic binding of clone 12 cells transfected with vector or plasmids expressing Y330A, Y336A, and del324–331 mutant forms of HAVCR1. (I) Luciferase expression in cell lysates of clone 12 cells transfected with Y330A, Y336A, or del324–331 mutant plasmids and infected with JFH1-Nanoluc. Data are from 4 independent experiments, each including 3 replicates, at 24 h postinfection. Horizontal bars represent the mean values. Asterisks represent significance values obtained using the Mann-Whitney test to compare the mean values for vector-transfected cells with the mean values for each of the other transfectants. NS, not significant; **, P < 0.001; ***, P < 0.0001. For FACS histograms, HAVCR1 expression on transfected cells is shown as filled dark-gray areas, and data for untransfected cells stained with anti-HAVCR1-1 MAb are shown as filled light-gray areas. RLU, relative light units; MFI, mean fluorescence intensity.

Article Snippet: To characterize receptor expression on the surface of Huh7.5 cells, we performed staining with the following antibodies: anti-CD81 mouse MAb (JS-81; BD Pharmingen, San Jose, CA); anti-SRB1 antibody (m1B9; BioLegend, San Diego, CA), anti-HAVCR1 phycoerythrin (PE)-labeled mouse MAb (REA384; Miltenyi Biotec, Inc., Auburn, CA), or purified HAVCR1-1 MAb, an antibody directed against the IgV domain of HAVCR1 that blocks binding of phospholipids.

Techniques: Expressing, Infection, Transfection, Plasmid Preparation, Binding Assay, Luciferase, Mutagenesis, MANN-WHITNEY, Staining, Fluorescence

Journal: Journal of Virology

Article Title: Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry

doi: 10.1128/JVI.01742-17

Figure Lengend Snippet: Schematic representation of the HAVCR1, mHavcr1, and chimeric proteins used in transient-transfection experiments. (A) Receptor domains and mutations in the human and mouse proteins. (B) Sequence alignment of HAVCR1, mouse Havcr1 (mHavcr1), and chimeric protein. Yellow highlighting denotes residues conserved between the molecules. The different receptor regions are defined by colored underlining as follows: red, IgV domain; blue, mucin domain; green, transmembrane domain; black, cytoplasmic domain. Mutated or deleted residues in HAVCR1 are shown in black boxes.

Article Snippet: To characterize receptor expression on the surface of Huh7.5 cells, we performed staining with the following antibodies: anti-CD81 mouse MAb (JS-81; BD Pharmingen, San Jose, CA); anti-SRB1 antibody (m1B9; BioLegend, San Diego, CA), anti-HAVCR1 phycoerythrin (PE)-labeled mouse MAb (REA384; Miltenyi Biotec, Inc., Auburn, CA), or purified HAVCR1-1 MAb, an antibody directed against the IgV domain of HAVCR1 that blocks binding of phospholipids.

Techniques: Transfection, Sequencing

Journal: Journal of Virology

Article Title: Determinants in the Ig Variable Domain of Human HAVCR1 (TIM-1) Are Required To Enhance Hepatitis C Virus Entry

doi: 10.1128/JVI.01742-17

Figure Lengend Snippet: The IgV domain of human HAVCR1 is responsible for facilitating HCV entry. (A) Expression of HAVCR1 or mHavcr1 on the surface of transfected clone 12 cells 48 h posttransfection. Receptor expression is shown as filled dark-gray areas, and data for stained, untransfected cells are shown as filled light-gray areas. (B) Apoptotic binding of clone 12 cells transfected with vector or plasmids expressing HAVCR1, mHavcr1, or chimeric human/mouse receptor. MFI, mean fluorescence intensity. (C) Luciferase expression in cell lysates of clone 12 cells transfected with expression plasmids and infected with JFH1-Nanoluc. RLU, relative light units. Data shown are from 5 experiments with transient transfection of the control vector and HAVCR1 and 3 independent experiments for mHavcr1, including 3 replicates each, at 24 h postinfection. Asterisks represent significance values obtained using the Mann-Whitney test to compare the data for vector-transfected cells with the data for each of the other transfectants or the parent cell line. NS, not significant; **, P < 0.001; ***, P < 0.0001.

Article Snippet: To characterize receptor expression on the surface of Huh7.5 cells, we performed staining with the following antibodies: anti-CD81 mouse MAb (JS-81; BD Pharmingen, San Jose, CA); anti-SRB1 antibody (m1B9; BioLegend, San Diego, CA), anti-HAVCR1 phycoerythrin (PE)-labeled mouse MAb (REA384; Miltenyi Biotec, Inc., Auburn, CA), or purified HAVCR1-1 MAb, an antibody directed against the IgV domain of HAVCR1 that blocks binding of phospholipids.

Techniques: Expressing, Transfection, Staining, Binding Assay, Plasmid Preparation, Fluorescence, Luciferase, Infection, Control, MANN-WHITNEY

Journal: Clinical and Experimental Immunology

Article Title: In‐utero coxsackievirus B4 infection of the mouse thymus

doi: 10.1111/cei.12893

Figure Lengend Snippet: Effects of CV‐B4 E2 in‐utero infection on T cell differentiation. Thymic T cell suspensions were prepared from thymuses, collected at different post‐inoculation (p.i.) times, on offspring from negative control and CV‐B4 E2‐inoculated [either at day 10 gestation (G) or 17G, by the intraperitoneal or the oral route] dams, stained with anti‐CD90, anti‐CD4 and anti‐CD8 antibodies and analysed by fluorescence activated cell sorter (FACS). Histograms represent the DN, DP, SP4 and SP8 populations among total thymocytes (CD90+ cells). Data are representative of four independent experiments with n = 3 thymuses per group and per time‐point in each. Results are expressed as mean percentages ± standard deviations; *P < 0·05 versus mock. DN = double‐negative CD4–CD8–; DP = double‐positive CD4+CD8+; SP4 = single‐positive CD4+CD8–; SP8: single‐positive CD4–CD8+.

Article Snippet: T cell staining and FACS analysis Three‐colour cell staining was performed with CD90/Thy‐1 rat anti‐mouse monoclonal antibody (mAb) (clone G7) phycoerythrin (PE) conjugate, CD4 rat anti‐mouse mAb (clone GK1·5) phycoerythrin‐cyanin (PE‐Cy) ® 5 conjugate and CD8a rat anti‐mouse mAb (clone 53–6·7) fluorescein (FITC) conjugate (all from Life Technologies, Paisley, UK) antibody mixture, and the preparation was incubated for 30 min at 4°C in the dark.

Techniques: In Utero, Infection, Cell Differentiation, Negative Control, Staining, Fluorescence

Journal: Clinical and Experimental Immunology

Article Title: In‐utero coxsackievirus B4 infection of the mouse thymus

doi: 10.1111/cei.12893

Figure Lengend Snippet: Effects of CV‐B4 E2 in‐utero infection on T cell differentiation. Thymic T cell suspensions were prepared from thymuses, collected at different post‐inoculation (p.i.) times, on offspring from negative control and CV‐B4 E2‐inoculated [either at day 10 gestation (G) or 17G, by the intraperitoneal or the oral route] dams, stained with anti‐CD90, anti‐CD4 and anti‐CD8 antibodies and analysed by fluorescence activated cell sorter (FACS). Histograms represent the DN, DP, SP4 and SP8 populations among total thymocytes (CD90+ cells). Data are representative of four independent experiments with n = 3 thymuses per group and per time‐point in each. Results are expressed as mean percentages ± standard deviations; *P < 0·05 versus mock. DN = double‐negative CD4–CD8–; DP = double‐positive CD4+CD8+; SP4 = single‐positive CD4+CD8–; SP8: single‐positive CD4–CD8+.

Article Snippet: Three‐colour cell staining was performed with CD90/Thy‐1 rat anti‐mouse monoclonal antibody (mAb) (clone G7) phycoerythrin (PE) conjugate, CD4 rat anti‐mouse mAb (clone GK1·5) phycoerythrin‐cyanin (PE‐Cy) ® 5 conjugate and CD8a rat anti‐mouse mAb (clone 53–6·7) fluorescein (FITC) conjugate (all from Life Technologies, Paisley, UK) antibody mixture, and the preparation was incubated for 30 min at 4°C in the dark.

Techniques: In Utero, Infection, Cell Differentiation, Negative Control, Staining, Fluorescence

Journal: Central-European Journal of Immunology

Article Title: An anti-DEC-205 monoclonal antibody stimulates binding of thymocytes to rat thymic dendritic cells and promotes apoptosis of thymocytes

doi: 10.5114/ceji.2014.47722

Figure Lengend Snippet: Immunoperoxidase staining of rat thymus, with HD83 mAb ( A-D ) and anti-MHC II mAb ( E-H ). Specific HD83 labeling is localized in the cortex ( A ) and less intense on some individual dendritic-like cells scattered in the medulla ( B ). Note stronger HD83 labeling of cortical epithelium, specifically at the cortico-medullary ( A ) and cortico-epithelial-free zones border ( C ). Perivascular epithelial cells (arrow) ( C ) and subcapsular epithelial cells (arrow) ( D ) are HD83 negative. Staining with OX-6 mAb shows the positivity of both cortical and medullary compartment ( E ). Note strong staining of medullary TEC and TDC ( F ), isolated macrophages and perivascular TEC (arrow) in epithelial-free zones ( G ), subcapsular epithelium (arrow) and cortical epithelium ( H ). Magnifications: A and E 20×; others 40×

Article Snippet: The following mouse anti-rat monoclonal antibodies (mAbs) were used: HD83, mouse anti-DEC-205 mAb cross-reactive with human, mouse, monkey and rat DEC-205 antigen, a kind gift from Dr. Chae Gyu Park (Severance Biomedical Science Institute, Yonsei University College of Medicine, Korea) [ ] and OX-6 mAb, (anti I-A) (Serotec).

Techniques: Immunoperoxidase Staining, Labeling, Negative Staining, Staining, Isolation

Journal: Central-European Journal of Immunology

Article Title: An anti-DEC-205 monoclonal antibody stimulates binding of thymocytes to rat thymic dendritic cells and promotes apoptosis of thymocytes

doi: 10.5114/ceji.2014.47722

Figure Lengend Snippet: Expression of DEC-205 on thymic stromal cells in vitro . TDC/thymocyte rosettes, established in vitro after 30 min of cell incubation, as described in Material and methods, were labeled with the immunoperoxidase staining. Note the positivity of TNC ( A ) and TDC ( C ) compared to Ir mAb controls ( B and D , respectively) (magnifications 40×). Thymocytes were negative ( C )

Article Snippet: The following mouse anti-rat monoclonal antibodies (mAbs) were used: HD83, mouse anti-DEC-205 mAb cross-reactive with human, mouse, monkey and rat DEC-205 antigen, a kind gift from Dr. Chae Gyu Park (Severance Biomedical Science Institute, Yonsei University College of Medicine, Korea) [ ] and OX-6 mAb, (anti I-A) (Serotec).

Techniques: Expressing, In Vitro, Incubation, Labeling, Immunoperoxidase Staining

Journal: Central-European Journal of Immunology

Article Title: An anti-DEC-205 monoclonal antibody stimulates binding of thymocytes to rat thymic dendritic cells and promotes apoptosis of thymocytes

doi: 10.5114/ceji.2014.47722

Figure Lengend Snippet: Effect of HD83 mAb on formation of TDC/thymocyte rosettes. The percentage of TDC forming rosettes after 30 min culture ( A ). The mean number of thymocytes per rosette ( B ). Microscopic analysis of TDC/thymocyte rosettes after 30 min incubation with TDC in the presence of HD83 or irrelevant mAb ( C ) (magnification: 40×), MGG staining. The results were given as mean ± SD (3 different experiments) after 200-500 cells with or without rosettes ( A ) or thymocytes in rosettes ( B ), were calculated in each experiment

Article Snippet: The following mouse anti-rat monoclonal antibodies (mAbs) were used: HD83, mouse anti-DEC-205 mAb cross-reactive with human, mouse, monkey and rat DEC-205 antigen, a kind gift from Dr. Chae Gyu Park (Severance Biomedical Science Institute, Yonsei University College of Medicine, Korea) [ ] and OX-6 mAb, (anti I-A) (Serotec).

Techniques: Incubation, Staining

Journal: Central-European Journal of Immunology

Article Title: An anti-DEC-205 monoclonal antibody stimulates binding of thymocytes to rat thymic dendritic cells and promotes apoptosis of thymocytes

doi: 10.5114/ceji.2014.47722

Figure Lengend Snippet: Effect of HD83 mAb on thymocyte apoptosis. Apoptosis of total thymocytes in culture with TDC after 6 hours of cultivation determined by morphological criteria ( A ), and flow cytometry staining with AnnexinV-FITC/PI ( B ). The percentage of apoptotic thymocytes within rosettes formed after 30 min compared to total thymocytes ( C ) determined by morphological criteria. The results were given as mean ± SD (3 different experiments) after 500 thymocytes ( A ), 5000 thymocytes ( B ) or 200-500 thymocytes within rosettes ( C ) were calculated in each experiment

Article Snippet: The following mouse anti-rat monoclonal antibodies (mAbs) were used: HD83, mouse anti-DEC-205 mAb cross-reactive with human, mouse, monkey and rat DEC-205 antigen, a kind gift from Dr. Chae Gyu Park (Severance Biomedical Science Institute, Yonsei University College of Medicine, Korea) [ ] and OX-6 mAb, (anti I-A) (Serotec).

Techniques: Flow Cytometry, Staining