Cells were mechanically homogenized, followed by staining for surface molecules using LIVE/DEAD fixable Aqua (Invitrogen, 1:1000 dilution) and anti-CD4 PE-Cy5 (RPA-T4, BD, 1:50 dilution), anti-CD3 PE-Cy7 (UCHT1, eBioscience, 1:50 dilution), anti-CD45RA V450 (H100, eBioscience, 1:100 dilution), anti-CXCR5 conjugated with Alexa Fluor 488 (RF8B2, eBioscience, 1:50 dilution), Alexa Fluor 647 (RF8B2, BD, 1:50 dilution), or biotin (RF8B2, BD Biosciencese, 1:50 dilution)/streptavidin APC-Cy7 (1:400 dilution), anti-PSGL-1 PE (KPL-1, BD, 1:50 dilution) or APC (FLEG, eBioscience, 1:50 dilution), anti-ICOS FITC (C398.4A, eBioscience, 1:50 dilution), anti-CCR7 FITC (150503, R&D Systems, 1:50 dilution), anti-CD62L FITC (DREG56, eBioscience, 1:50 dilution), anti-CXCR4 PE-Cy5 (12G5, eBioscience, 1:50 dilution), anti-CD200 APC (OX104, eBioscience, 1:50 dilution), anti-OX40 PE-Cy5 (ACT35, BD, 1:50 dilution), anti-PD-1 PE-Cy7 (EH12.1, BD, 1:50 dilution), anti-CXCR3 BV421(1C6/CXCR3, BD, 1:50), anti-IL-2RA PE (M-A251, BD, 1:25), anti-CD19 APC-Cy7 (SJ25C1, eBioscience, 1:50 dilution), anti-IgD PE-Cy7 (IA6-2, BD, 1:50 dilution), anti-CD38 V450 (HIT2, BD, 1:50 dilution), and anti-IL-10R PE (3F9, Biolegend, 1:50 dilution).
Anti cxcr3 bv421
Manufactured by BD
Sourced in United States
Anti-CXCR3 BV421 is a fluorochrome-conjugated antibody that binds to the CXCR3 chemokine receptor. CXCR3 is expressed on the surface of certain types of immune cells and plays a role in their trafficking and activation. The BV421 fluorochrome allows for the detection and analysis of CXCR3-expressing cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd3 pe cy7, by Thermo Fisher Scientific (1 mentions)
Anti igd pe cy7, by BD (1 mentions)
Anti cd38 v450, by BD (1 mentions)
Anti cd19 apc cy7, by Thermo Fisher Scientific (1 mentions)
Pe cy5 anti cd4, by BD (1 mentions)
Live dead fixable aqua, by Thermo Fisher Scientific (1 mentions)
Anti pd 1 pe cy7, by BD (1 mentions)
Rpa t4, by BD (1 mentions)
Flow cytometry buffer, by Thermo Fisher Scientific (1 mentions)
Lsr 2 cytometer, by BD (1 mentions)
Facsdiva software, by BD (1 mentions)
Cd69 pe, by Thermo Fisher Scientific (1 mentions)
Cd8 af700, by BD (1 mentions)
Tbet pe, by Thermo Fisher Scientific (1 mentions)
Il 17a pecy7, by BD (1 mentions)
Anti ifn γ pe, by BD (1 mentions)
Anti cd19 fitc, by BD (1 mentions)
Anti cd4 fitc, by BD (1 mentions)
Cd4 fitc, by BD (1 mentions)
Anti foxp3 pe, by Thermo Fisher Scientific (1 mentions)
3 protocols using anti cxcr3 bv421
1
Multiparametric Flow Cytometry Immunophenotyping
2
Quantification of Tumor-Infiltrating Th17 Cells
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Xenograft tumors from mice groups (4 per group) described in the previous section were harvested 12 days after tumor formation induction and then dissociated in RPMI with the Lysates were centrifuged and cells were stained with an antibody cocktail containing anti-CXCR3-BV421 (CXCR3-173, BD Biosciences), anti-CD4-VioBright FITC (REA604, Miltenyi Biotech), anti-CD45RA-APC-Vio770 (30F11, Miltenyi Biotech) and anti-CCR6-PE (REA277, Miltenyi Biotech). Tumor infiltrating Th17 cells were identified as CD4 + , CD45RA -, CCR6 + , CXCR3 -. After surface staining, 2 Ml of red blood cells lysis solution (BD Biosciences) was added for 10 minutes, centrifuged (400 x g, 5 minutes) and then resuspended in flow cytometry buffer (eBioscience). All events were acquired by a BD LSR-II cytometer equipped with BD FACSDiva software (BD Biosciences), and data were analyzed using FlowJo software (Tree Star). The gating strategy is illustrated in Figure 5A.
3
Comprehensive Immune Cell Analysis in Murine Transplantation
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The spleen and bone marrow of the model mice and healthy recipient control mice
were harvested at the 6th or 16th week after BMT. Single cell suspensions from
spleens and bone marrows were labeled with anti-CD3 PE, -CD4 FITC, -CD8 AF700
(BD Pharmingen, San Diego, CA, USA) for T cells, anti-CD19 FITC (BD) for B
cells, anti-CXCR3 BV421 and anti-CD80 FITC for immune molecular examination,
anti-CD4 FITC (BD) and anti-Foxp3 PE (eBioscience, San Diego, CA, USA) for
regulatory T cells (Treg), anti-IFN-γ PE and -IL-17A PE-cy7 (BD) for Th1/Th17
subsets, respectively. Before detecting the secretion of IFN-γ and IL-17A in T
cells, cell suspensions were activated with anti-mouse CD3/CD28 microbeads for 3
hours. The spleen mononuclear cells of mice transplanted with A20 lymphoma cells
were stained with anti-B220 PE and -H-2Kb BV421 (eBioscience) to
examine the ratio of tumor cells. Activated human PBMCs with anti- human
CD3/CD28 beads were stained with anti-human CD3 FITC and -CD69 PE for T cell
activation analysis or with anti-human CD4 PE-cy7, -T-bet PE, -Foxp3 AF647,
-GATA3 APC and -ROR?t PE (eBioscience) for T cell differentiation analysis after
incubation with SHR0302 (1 μM) for 24 h. Fixable viability dye or DAPI (BD) were
used to distinguish live cells from dead cells.
were harvested at the 6th or 16th week after BMT. Single cell suspensions from
spleens and bone marrows were labeled with anti-CD3 PE, -CD4 FITC, -CD8 AF700
(BD Pharmingen, San Diego, CA, USA) for T cells, anti-CD19 FITC (BD) for B
cells, anti-CXCR3 BV421 and anti-CD80 FITC for immune molecular examination,
anti-CD4 FITC (BD) and anti-Foxp3 PE (eBioscience, San Diego, CA, USA) for
regulatory T cells (Treg), anti-IFN-γ PE and -IL-17A PE-cy7 (BD) for Th1/Th17
subsets, respectively. Before detecting the secretion of IFN-γ and IL-17A in T
cells, cell suspensions were activated with anti-mouse CD3/CD28 microbeads for 3
hours. The spleen mononuclear cells of mice transplanted with A20 lymphoma cells
were stained with anti-B220 PE and -H-2Kb BV421 (eBioscience) to
examine the ratio of tumor cells. Activated human PBMCs with anti- human
CD3/CD28 beads were stained with anti-human CD3 FITC and -CD69 PE for T cell
activation analysis or with anti-human CD4 PE-cy7, -T-bet PE, -Foxp3 AF647,
-GATA3 APC and -ROR?t PE (eBioscience) for T cell differentiation analysis after
incubation with SHR0302 (1 μM) for 24 h. Fixable viability dye or DAPI (BD) were
used to distinguish live cells from dead cells.
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