Cxcr5 apc

Manufactured by BioLegend

Sourced in United States

CXCR5-APC is a fluorochrome-conjugated antibody that binds to the CXCR5 chemokine receptor. CXCR5 is expressed on B cells and a subset of T cells and plays a role in homing to B cell follicles. The APC fluorochrome allows for detection of CXCR5-positive cells by flow cytometry.

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Lab products found in correlation

Pd 1 pe, by BioLegend (2 mentions) Pd1 pe cy7, by BioLegend (2 mentions) Cd3 fitc, by BioLegend (1 mentions) Cd19 pe, by BioLegend (1 mentions) Percp cy5.5 cd4, by BioLegend (1 mentions) Facscalibur, by BD (1 mentions) Cd4 pe, by Thermo Fisher Scientific (1 mentions) Pe cy7 pd 1, by BioLegend (1 mentions) Cyan adp analyzer, by Beckman Coulter (1 mentions) Cd4 pe, by BioLegend (1 mentions) Cxcr4 percp efluor710, by Thermo Fisher Scientific (1 mentions) Cd69 pe, by BioLegend (1 mentions) Canto 2, by BD (1 mentions) Lsrii cytometer, by BD (1 mentions) Cd80 pe cy5, by Thermo Fisher Scientific (1 mentions) Cd19 percp cy5, by Thermo Fisher Scientific (1 mentions) H3n2 ha, by Sino Biological (1 mentions) H1n1 ha, by Sino Biological (1 mentions) H5n1 ha, by Sino Biological (1 mentions) Cd138 pe, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Anti-CXCR5-APC (12 citations)

8 protocols using cxcr5 apc

Phenotyping of circulating T follicular helper cells

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Peripheral blood mononuclear cells were isolated by density gradient centrifugation as previously reported (17 (link)). After washed twice with phosphate-buffered saline (PBS), PBMCs were incubated with the following fluorochrome-conjugated monoclonal Abs: FITC-CD3, PerCP-Cy5.5-CD4, APC-CXCR5, PE-PD-1, PE-CD19, and relevant isotype controls (Biolegend, San Diego, CA, USA). After staining at 4°C for 30 min, PBMCs were washed twice with PBS containing 2% fetal bovine serum and then measured on a BD FACS Calibur instrument. cTfh cells were defined as CD3⁺CD4⁺CXCR5⁺PD-1⁺. Data were analyzed using the FlowJo 7.6 software.

Zhao C., Li H.Z., Zhao D.D., Ma C., Wu F., Bai Y.N., Zhang M., Li Z.Y, & Guo J. (2017). Increased Circulating T Follicular Helper Cells Are Inhibited by Rituximab in Neuromyelitis Optica Spectrum Disorder. Frontiers in Neurology, 8, 104.

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Immunophenotyping of PBMCs by Flow Cytometry

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PBMCs were immunostained with fluorescein isothiocyanate (FITC)-ICOS, phycoerythrin (PE)-CD4 (eBioscience, USA), and APC-CXCR5, PE-Cy7-PD-1 (BioLegend, USA) monoclonal antibodies according to the manufacturers' protocols. The stained cells were then analyzed using a CyAn ADP analyzer (Beckman Coulter) and data were analyzed with FlowJo software.

Zhang Y., Jiang Y., Wang Y., Liu H., Shen Y., Yuan Z., Hu Y., Xu Y, & Cao J. (2015). Higher Frequency of Circulating PD-1high CXCR5+CD4+ Tfh Cells in Patients with Chronic Schistosomiasis. International Journal of Biological Sciences, 11(9), 1049-1055.

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Multicolor Flow Cytometry Analysis of Lymphocytes

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Lymphocytes isolated from C57BL/6J popliteal lymph nodes by manual dissociation through a 35 μm filter were stained extracellularly by incubation with α-mouse B220-APC, -PerCPCy5.5, or -AF488 (BioLegend, San Diego, CA), IgM-eFluor® 450 (eBioscience), CD3ε-APC/Cy7 or -APCeFluor780 (eBioscience), S1PR1-APC (R&D Systems), CD4⁺-PE (BioLegend), CD8⁺-AF488, -APC or -PacBlue (BioLegend), BrdU-FITC (Invitrogen), CD69-PE (BioLegend), CXCR4-PerCP-eFluor® 710 (eBioscience), CXCR5-APC (BioLegend), and CCR7-e450 (eBioscience) conjugated antibodies. Stained cells were fixed and acquired with a Canto II or LSR II cytometer (BD Biosciences) and analyzed using FlowJo software (Tree Star). At least 50,000 lymphocytes were analyzed for each sample. Lymphocytes were identified by forward/side scatter profiles as depicted in Fig. 1.

Habenicht L.M., Albershardt T.C., Iritani B.M, & Ruddell A. (2016). Distinct mechanisms of B and T lymphocyte accumulation generate tumor-draining lymph node hypertrophy. Oncoimmunology, 5(8), e1204505.

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Binding Interactions of Influenza Hemagglutinin Subtypes

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BSA and streptavidin were from Sigma-Aldrich. PBS was from Gibco. TT was supplied by the Statens Serum Institut. All recombinant influenza HA subtypes were from Sino Biological: H1N1 HA (A/California/04/2009; H1), H2N2 HA (A/Japan/305/1957; H2), H5N1 HA (A/Vietnam/1194/2004; H5), H3N2 HA (A/Perth/16/2009; H3) and H7N9 HA (A/Shanghai/1/2013; H7). H1N1 HA (A/New Caledonia/20/1999; H1 wt), the HA delta stem mutants H1 I45R/T49R and H1 45glyc, and the human monoclonal antibodies CR6261 and CH65 were provided by D. Lingwood. HIV-1 92BR-gp120 was provided by D.R. Burton (Scripps Institute, San Diego, CA). Human CpG ODN 2006 was from InvivoGen. IL-15 and IL-6 were from PeproTech. Antibodies used were as follows: goat anti–human IgG (FcΥ specific; Jackson ImmunoResearch Laboratories, Inc.); anti–human CD27-APC, anti–human CD38-FITC, CD138-PE, CD19-Percp Cy5.5, CD20-APC, CD80-PE-Cy5, HLA-DR-PE-Cy5, and anti–human κ-Ig biotin (eBioscience); anti-human CD86-Bv510, CCR10-PE, CD62L, CXCR4-PE-Cy5, CXCR5-APC, and anti–human λ-Ig biotin (BioLegend); and anti–human CD43-PE and anti–human IgD (Miltenyi Biotec).

Sanjuan Nandin I., Fong C., Deantonio C., Torreno-Pina J.A., Pecetta S., Maldonado P., Gasparrini F., Ordovas-Montanes J., Kazer S.W., Kjaer S., Borley D.W., Nair U., Coleman J.A., Lingwood D., Shalek A.K., Meffre E., Poignard P., Burton D.R, & Batista F.D. (2017). Novel in vitro booster vaccination to rapidly generate antigen-specific human monoclonal antibodies. The Journal of Experimental Medicine, 214(8), 2471-2490.

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Quantifying T Cell Subsets by Flow Cytometry

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To quantify CD4⁺ICOS⁺CD38⁺ and CD8⁺ICOS⁺CD38⁺ T cells, PBMCs were first resuspended with Human TruStain Fcx (Biolegend) for 10 min at room temperature and then stained with the following antibodies in FACS buffer (PBS + 2% fetal bovine serum): CD8a e450 (Invitrogen 48-0086-42, 1:200) ICOS BV605 (Biolegend 313538, 1:50), CCR7 PE (Biolegend 353204, 1:200), CD38 PerCP (Biolegend 303520, 1:100), CD4 PECy7 (Biolegend 357410, 1:100), and CD3 APCCy7 (Biolegend 300318, 1:200). Tfh markers consisted of CXCR5 APC (Biolegend 356907, 1:200) and PD-1 PE (Biolegend 135205, 1:100). CD71 APC (Biolegend 334108, 1:100) was also measured on sorted cells. Cells were analyzed on a Miltenyi MACSQuant16 Analyzer with single-stain control PBMC samples used for compensation conducted in FlowJo v10.6.2.

Kramer K.J., Wilfong E.M., Voss K., Barone S.M., Shiakolas A.R., Raju N., Roe C.E., Suryadevara N., Walker L.M., Wall S.C., Paulo A., Schaefer S., Dahunsi D., Westlake C.S., Crowe JE J.r., Carnahan R.H., Rathmell J.C., Bonami R.H., Georgiev I.S, & Irish J.M. (2022). Single-cell profiling of the antigen-specific response to BNT162b2 SARS-CoV-2 RNA vaccine. Nature Communications, 13, 3466.

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Characterizing Tfh and B Cells by Flow Cytometry

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Peripheral blood mononuclear cells (PBMCs) were collected and isolated by Ficoll-density gradient centrifugation. PBMCs were immuno-stained for 30 min with the following antibodies, Tfh cells: CD4-PE, ICOS-FITC (eBioscience, San Diego, CA,USA), CXCR5-APC and PD-1 PE-Cy7 (BioLegend, San Diego, CA, USA); circulating B cells: CD4-FITC, CD19-APC and CD27-PE. The stained cells were then assessed using a CyAn ADP cytometer (Beckman Coulter, Lambertville, NJ, USA) and data were analyzed with FlowJo software.

Zhang Y., Wang Y., Jiang Y., Pan W., Liu H., Yin J., Shen Y, & Cao J. (2016). T follicular helper cells in patients with acute schistosomiasis. Parasites & Vectors, 9, 321.

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Profiling T Cell Subsets in BAL Samples

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A panel of antibodies was used to differentiate CD4⁺ T cell subsets from BAL samples of infected vaccinated mice. BAL cells were stimulated with NP₅₅ and HA₁₄₀ peptide, as previously described [11 (link)], for cytokine production following 2 h peptide stimulation in the presence of Brefeldin A for a further 4 h, for a total 6 h stimulation. The FcR was blocked prior to immune cell staining, 20 min at room temperature (anti-CD16/CD32, BD Bioscience). Cells were stained for surface markers for anti-mouse CD3-BV510, CD4-APCCy7, CD25-PerCPCy5.5, CXCR5-APC and PD1-BV605 (all Biolegend). Samples were then fixed with FoxP3 Fix/Perm buffer (eBioscience), and intracellular stain for IFN-γ-FITC, RORγt-PE, IL-17A-AF700, FoxP3-Pacific Blue, Bcl6-PETexas and IL-4-PECy7. Cells were acquired by flow cytometry on a LSR Fortessa and analysed by FlowJo software. Live lymphocyte gating, followed by CD3⁺ CD4⁺ to determine: Th2 (IL-4⁺), Th1 (IFN-γ⁺), Th17 (IL-17A⁺), Treg (CD25⁺ FoxP3⁺) and Tfh cells (CXCR5⁺).

Valkenburg S.A., Li O.T., Li A., Bull M., Waldmann T.A., Perera L.P., Peiris M, & Poon L.L. (2018). Protection by universal influenza vaccine is mediated by memory CD4 T cells. Vaccine, 36(29), 4198-4206.

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Multiparameter Flow Cytometry Analysis

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Flow cytometry was performed using the following anti-mouse antibodies: PD1-FITC, PD1-PE/cy7, LAG3-PE, CD4-PerCP/Cy5.5, KLRG1-APC, CD8-PE/cy7, CD8-APC/cy7, CD28-APC, T-bet-APC, CXCR5-APC (Biolegend, Cal., US); CD3-FITC,CD3-PerCP/Cy5.5 CD57-FITC, CD8-PE, CTLA4-PE, Tim3-APC, CD56-FITC, CD56-APC/cy7, CD19-APC/cy7, CD45-BV421 (BD Biosciences, NJ, US); Eomes-FITC (eBioscience, Cal., US). Flow cytometry analysis was performed on FACS Canto II (BD Biosciences, NJ, US) and analyzed with FlowJo software.

Liu H., Xu J., Wang L., Mao W., Wang B., Liang Z., Zhang Y., Shi Y., Tang B., Shen Y., Dong Y., Ren H., & Cen X. (2020). T Cell Exhaustion and Senescence in Epstein-Barr Virus-Associated Lymphoma Patients.

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