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Anti cxcr5 bv421

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Anti-CXCR5-BV421 is a fluorochrome-conjugated antibody that binds to the CXCR5 chemokine receptor. CXCR5 is a G protein-coupled receptor that plays a key role in the homing and migration of B cells. The BV421 fluorochrome allows for detection and analysis of CXCR5-expressing cells using flow cytometry.

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Lab products found in correlation

Anti cd4 bv650, by BioLegend (2 mentions) Cd69 fitc, by BD (1 mentions) Anti cd8 af700, by BD (1 mentions) Anti cd44 bv510, by BD (1 mentions) Anti cd62l pe cy7, by BD (1 mentions) Anti pd 1 bv711, by BioLegend (1 mentions) Pan t cell isolation kit 2, by Miltenyi Biotec (1 mentions) Zombie aqua viability dye, by BioLegend (1 mentions) Anti cd14 bv510 clone m5e2, by BioLegend (1 mentions) Anti pe and anti apc magnetic beads, by Miltenyi Biotec (1 mentions) Clone j252d4, by BioLegend (1 mentions) Lsrfortessa x 20, by BD (1 mentions) Transcription factor fix perm buffer, by Thermo Fisher Scientific (1 mentions) Anti pd 1 pe dazzle 594, by BioLegend (1 mentions) Fixable viability dye efluor 455uv, by Thermo Fisher Scientific (1 mentions) Fortessa analyzer, by BD (1 mentions) Anti cd3 af700, by BioLegend (1 mentions) Anti ccr2 pe, by BioLegend (1 mentions) Anti cd4 pe cy7, by BD (1 mentions) Aurora analyzer, by Cytek (1 mentions)

4 protocols using anti cxcr5 bv421

1

Tracking Antigen-Specific T Cell Responses

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C57Bl/6 mice (Thy1.2+) were challenged with DENV1, JEV, and YFV (five mice per group) by intraperitoneal injection with 1 × 106 PFU or injected with saline. Five weeks after infection, the spleens were collected and the total T cells were purified by magnetic column using the Pan T Cell Isolation Kit II (mouse; MACS Miltenyi Biotec). C57Bl/6 mice (Thy1.1+) received an adoptive transfer by tail vein injection of 1 × 106 total T cells per mouse, with either T cells purified from mice exposed to DENV1, YFV, JEV, or saline. The mice were challenged with DENV1 24 hours after transfer, and after 6 days, popliteal LNs were collected and prepared as single-cell suspensions for flow cytometry. The cells were stained using the following primary conjugated antibodies: anti-Thy1.2–PerCP Cy5.5 (BioLegend), anti-CD4–BV650, anti-CD8–AF700, anti-CD44–BV510, anti-CD62L–PE-Cy7, CD69-FITC (all from BD Biosciences), anti-Bcl6–AF647 (BioLegend), anti–PD-1–BV711 (BioLegend), and anti-CXCR5–BV421 (BioLegend).
Saron W.A., Rathore A.P., Ting L., Ooi E.E., Low J., Abraham S.N, & St. John A.L. (2018). Flavivirus serocomplex cross-reactive immunity is protective by activating heterologous memory CD4 T cells. Science Advances, 4(7), eaar4297.
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2

SARS-CoV-2 Antigen-Specific T Cell Profiling

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Cryopreserved samples containing 1-10 × 107 PBMC from DRB1*07:01 (DR7)+ convalescent subjects or pre-pandemic negative controls were thawed, washed twice with PBS containing 2% FBS, and resuspended in either complete EHAA or RPMI 1640 with 5% FBS. Staining with a pre-mixed cocktail of DR7:p-streptavidin-PE, DR7:p-streptavidin-PE-Cy7, and DR7:p-streptavidin-APC tetramers at a final concentration of 10 nM for each reagent was for 1 hour at room temperature. Anti-PE and anti-APC magnetic beads (Miltenyi Biotec) were then added, and bead-bound cells were enriched as previously described (22). Cells in the bound and unbound fractions were stained with Zombie Aqua viability dye (BioLegend) and a surface marker antibody panel consisting of anti-CD20BV510 (clone 2H7, BioLegend), anti-CD14BV510 (clone M5E2, BioLegend), anti-CD3AF700 (clone UCHT1, BioLegend), anti-CD4BV605 (clone OKT4, BioLegend), anti-CD8BUV395 (clone RPA-T8, BioLegend), anti-CD45ROAPC/Cy7 (clone UCHL1, BioLegend), anti-CXCR3PE/Dazzle594 (clone G025H7, BioLegend), anti-CXCR5BV421 (clone J252D4, BioLegend), anti-PD-1BV785 (clone EH12.2H7, BioLegend), and anti-CCR4PerCP/Cy5.5 (clone L291H4, BioLegend) for 30 min at room temperature. Data were acquired on an LSR Fortessa X-20 (BD) flow cytometer and analyzed with FlowJo software (BD).
Nelson R.W., Chen Y., Venezia O.L., Majerus R.M., Shin D.S., Carrington M.N., Yu X.G., Wesemann D.R., Moon J.J, & Luster A.D. (2022). SARS-CoV-2 epitope-specific CD4+ memory T cell responses across COVID-19 disease severity and antibody durability. Science Immunology.
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3

Multiparametric Phenotyping of Immune Cells

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Synovial tissue and synovial fluid cells were thawed, washed twice in PBS, and incubated with Fixable Viability Dye eFluor 455UV (eBioscience) for 30 minutes. Cells were then washed in PBS/1%BSA and stained with antibodies against surface markers anti-CD3-AF700, anti-CD4-BV650, anti-CCR2-PE, anti-CXCR5-BV421, anti-PD-1-PE/Dazzle 594 (all Biolegend) for 30 minutes. Cells were washed once and incubated with eBioscience Transcription Factor Fix/Perm Buffer. Cells were washed in PBS/1%BSA/0.3% saponin and incubated in intracellular antibodies anti-MAF-PerCP-eFluor710 (sym0F1, eBioscience), anti-Bcl6-APC (BCL-UP, eBioscience), and anti-Blimp-1-AF488 (646702, R&D Systems) at 1:20 dilutions for 4 hours. Cells were washed once, filtered, and data acquired on a BD Fortessa analyzer. Intracellular detection of FoxP3 and CTLA-4 were performed by the same method on magnetic-bead purified blood CD4+ T cells using the indicated surface markers.
Rao D.A., Gurish M.F., Marshall J.L., Slowikowski K., Fonseka C.Y., Liu Y., Donlin L.T., Henderson L.A., Wei K., Mizoguchi F., Teslovich N.C., Weinblatt M.E., Massarotti E.M., Coblyn J.S., Helfgott S.M., Lee Y.C., Todd D.J., Bykerk V.P., Goodman S.M., Pernis A.B., Ivashkiv L.B., Karlson E.W., Nigrovic P.A., Filer A., Buckley C.D., Lederer J.A., Raychaudhuri S, & Brenner M.B. (2017). Pathologically expanded peripheral T helper cell subset drives B cells in rheumatoid arthritis. Nature, 542(7639), 110-114.
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4

Exercise-Induced Changes in Lymphocyte Phenotypes

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Peripheral blood samples were obtained by venous puncture before exercise (baseline period, T0), at exercise exhaustion (post-exercise period, T1), and after resting one hour (resting period, T2). Peripheral blood mononuclear cells (PBMCs) were isolated from blood samples collected in EDTA tubes by Ficoll-Hypaque density gradient centrifugation, followed by cryopreservation in fetal calf serum with 10 % dimethyl sulfoxide until time of analysis. Flow cytometry of samples was analyzed using a previously published methodology (Martínez-Rodríguez et al., 2011) , staining samples with the following monoclonal antibodies: anti-CD3-PerCP, anti-CD56-BV510, anti-CD8-Alexa Fluor 700, anti-CD4-PECy7 (BD Pharmingen), and anti-CD19-PECy7 (eBiosciences). NK cells were defined as CD3-CD56+ cells, identifying CD56 dim and CD56 bright NK cell subsets based on the staining intensity of the specific CD56 monoclonal antibody; T cells and B cells were defined as CD3+ and CD3-CD19+ lymphocytes, respectively. Chemokine receptors expression by lymphocytes were assessed using the following monoclonal antibodies: anti-CXCR5-BV421, anti-CX3CR1-BV650, anti-CXCR3-BV711, anti-CCR7-AF647, anti-CCR1-PE, and anti-CCR5-BV785 (Biolegend). Samples were acquired in full spectrum Cytek Aurora Analyzer (Cytek Biosciences, Fremont, USA), analyzing data using FlowJo software (tree Star, Oregon, USA).
Munteis E., Vera A., Llop M., Moreira A., Oviedo G.R., Javierre C, & Martínez-Rodríguez J.E. (2024). Decreased exercise-induced natural killer cell redistribution in multiple sclerosis. Multiple sclerosis and related disorders, 87.
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