Human MNCs were isolated from venous blood samples by Ficoll-Paque PLUS (GE Healthcare, Chicago, IL, USA), and urine cell pellets were obtained from fresh urine specimens by centrifugation and washing procedures. Mouse spleens were homogenized by using syringe plunger and mesh strainer to collect splenocytes. Human MNCs were incubated with PE/Cy5 anti-human CD4 (BD Pharmingen, San Diego, CA, USA) and FITC anti-human CD14 (BD Pharmingen) antibodies, and mouse splenocytes were incubated with PE/Cy5 anti-mouse CD4 (BD Pharmingen) and FITC anti-mouse CD19 (BD Pharmingen) antibodies. These cells were sorted by Moflo XDP Cell Sorter (Beckman Coulter, Mountain View, CA, USA) to obtain human CD4+ and CD14+ cells and mouse CD4+ and CD19+ cells with the purity up to 95%.
Fitc anti mouse cd19
Manufactured by BD
Sourced in United States
FITC anti-mouse CD19 is a fluorescently labeled antibody that binds to the CD19 surface antigen expressed on mouse B cells. It is a tool used in flow cytometry experiments to identify and study B cell populations.
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Lab products found in correlation
Anti human cd14 fitc, by BD (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Moflo xdp cell sorter, by Beckman Coulter (1 mentions)
Anti mouse igg hrp, by Cell Signaling Technology (1 mentions)
Recombinant mouse il 4, by Miltenyi Biotec (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
Anti mouse cd40, by BD (1 mentions)
Lps escherichia coli o26 b6, by Merck Group (1 mentions)
Anti mouse igm f ab 2, by Jackson ImmunoResearch (1 mentions)
Pe anti mouse cd40, by BD (1 mentions)
April, by BD (1 mentions)
Fitc hamster anti mouse cd3e, by BD (1 mentions)
Rat anti mouse cd8a percp, by BD (1 mentions)
Pe rat anti mouse cd4, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Anti mouse cd4, by BD (1 mentions)
Anti mouse cd3 fitc, by BD (1 mentions)
Streptavidin apc cy7, by BD (1 mentions)
Alexa fluor 700 anti mouse igd, by BioLegend (1 mentions)
Apc cy7 anti mouse cd8a, by BioLegend (1 mentions)
5 protocols using fitc anti mouse cd19
1
Isolation and Sorting of Human and Mouse Immune Cells
2
Murine C5a: A Comprehensive Immunological Assay
3
Murine Splenic Lymphocyte Characterization
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At 21 and 42 days of age, splenic samples of eight mice in each group were taken to determine the percentages of CD3+, CD4+, CD8+ T lymphocyte and CD 19+ B lymphocyte by FCM.
Each spleen was cut into pieces and then filtered through nylon gauze as splenic single-cell suspension. The suspension was centrifuged at 200 × g for 5 min. The supernatant was discarded and lymphocytes were collected. The cell concentration was determined by using the normal counting method of blood cells and then diluted to 1.0 × 106 cells/mL with phosphate-buffered saline (PBS). A total of 100 μL cell suspensions was transferred to another centrifuge tube. The cells were respectively stained with 10 μL hamster anti-mouse CD3e-FITC (BD, Cat No: 553062), rat anti-mouse CD4-PE (BD, Cat No: 557308), rat anti-mouse CD8a-PerCP (BD, Cat No: 553036) and FITC anti-mouse CD19+(BD, Cat No: 553785) for 30 min at RT, and then 2 mL PBS was added and centrifuged at 200 × g for 5 min. The supernatant was discarded. Cells were resuspended in 0.5 mL PBS and determined by BD FACS Calibur flow cytometer.
Each spleen was cut into pieces and then filtered through nylon gauze as splenic single-cell suspension. The suspension was centrifuged at 200 × g for 5 min. The supernatant was discarded and lymphocytes were collected. The cell concentration was determined by using the normal counting method of blood cells and then diluted to 1.0 × 106 cells/mL with phosphate-buffered saline (PBS). A total of 100 μL cell suspensions was transferred to another centrifuge tube. The cells were respectively stained with 10 μL hamster anti-mouse CD3e-FITC (BD, Cat No: 553062), rat anti-mouse CD4-PE (BD, Cat No: 557308), rat anti-mouse CD8a-PerCP (BD, Cat No: 553036) and FITC anti-mouse CD19+(BD, Cat No: 553785) for 30 min at RT, and then 2 mL PBS was added and centrifuged at 200 × g for 5 min. The supernatant was discarded. Cells were resuspended in 0.5 mL PBS and determined by BD FACS Calibur flow cytometer.
4
Lymphocyte Subsets and Cell Cycle Analysis
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The percentages of cultured splenic lymphocyte subsets were measured by flow cytometer, using monoclonal fluorescein isothiocyanate (FITC) anti-mouse CD3+, phycoerythrin (PE) anti-mouse CD4+, PerCp anti-mouse CD8+ and FITC anti-mouse CD19+ (BD Biosciences, San Jose, CA, USA) as monoclonal antibodies labeled, respectively. The results were analyzed using the Cell Quest computer program.
The cell cycle was measured by flow cytometer. Cells were incubated for 30 min at room temperature in the dark with 0.15% Triton X-100 and propidium iodide (PI). The results were analyzed by the use of the Mod Fit LT for Mac V3.0 computer program.
The cell cycle was measured by flow cytometer. Cells were incubated for 30 min at room temperature in the dark with 0.15% Triton X-100 and propidium iodide (PI). The results were analyzed by the use of the Mod Fit LT for Mac V3.0 computer program.
5
Comprehensive Lung Cell Immunophenotyping
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Single lung-cell suspensions were incubated using a Zombie Aqua™ Fixable Viability Kit (BioLegend, San Diego, CA, USA) at RT for 30 min to discriminate dead cells for analysis. Cells were washed with DPBS containing 1% FBS, blocked with anti-CD16/CD32 (clone 2.4G2, ATCC HB-197), and then stained with FITC anti-mouse CD19 (BD Biosciences), PE anti-mouse CD69 (BioLegend), PerCP-Cy5.5 anti-mouse CD3e (BD Biosciences), PE-Cy7 anti-mouse CD103 (BioLegend), APC anti-mouse CD44 (BioLegend), APC-Cy7 anti-mouse CD8a (BioLegend), Alexa Fluor 700 anti-mouse CD4 (BD Biosciences), or BV421 anti-mouse CD45.2 (BD Biosciences) antibodies for 30 min at 4 °C. For B-cell analysis, cells were stained with FITC anti-mouse IgA (Southern Biotech), PE anti-mouse CD19 (BioLegend), PerCP-Cy5.5 anti-mouse CD38 (BioLegend), PE-Cy7 anti-mouse CD3e (BD Biosciences), biotin anti-mouse CD45.2 (eBioscience), Alexa Fluor 700 anti-mouse IgD (BioLegend), Brilliant Violet™ 421 anti-mouse CD80 (BioLegend), or streptavidin APC-Cy7 (BD Biosciences) antibodies for 30 min at 4 °C. After staining, cells were washed with DPBS containing 1% FBS and analyzed on a FACS Fortessa (BD Biosciences) using FlowJo Software Version 10 (Tree Star, Ashland, OR, USA). The gating strategy is presented in Supplementary Fig. 1 .
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