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Anti cxcr5

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Anti-CXCR5 is a monoclonal antibody that specifically recognizes the CXCR5 chemokine receptor. CXCR5 is involved in the homing and migration of B cells and a subset of T cells. This antibody can be used to detect and study CXCR5-expressing cells.

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Lab products found in correlation

Anti cd11c, by BioLegend (4 mentions) Anti cd3, by BioLegend (3 mentions) Foxp3 fix perm kit, by Thermo Fisher Scientific (2 mentions) Anti cxcr3, by BioLegend (2 mentions) Anti igm, by BioLegend (2 mentions) Anti igg, by BD (2 mentions) Anti cd38, by BD (2 mentions) Anti igd, by Thermo Fisher Scientific (2 mentions) Unlabeled mouse igg, by BioLegend (2 mentions) Anti cd27, by Thermo Fisher Scientific (2 mentions) Anti ccr5, by Thermo Fisher Scientific (2 mentions) Lsr fortessa cytometer, by BD (2 mentions) Anti iga, by Miltenyi Biotec (2 mentions) Anti cd20, by BioLegend (2 mentions) Anti ige, by BioLegend (2 mentions) Anti icam 1, by BioLegend (2 mentions) T bet, by BioLegend (2 mentions) Anti cd19, by BioLegend (2 mentions) Live dead aqua, by Thermo Fisher Scientific (2 mentions) Anti pd 1, by BioLegend (2 mentions)

5 protocols using anti cxcr5

1

Comprehensive Immunophenotyping of LNMC and PBMC

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Two million of LNMC or PBMC were stained for flow cytometry as previously described to analyze the expression of surface and intracellular markers as well as cellular composition using the 3-laser FACS Fortessa (Becton Dickinson; San Jose, USA)(16 (link)). Antibodies used in these analyses were purchased from either BD or Biolegend and included anti-CD16 (clone 3G8), anti-CD11b (clone M1/70), anti-CD1c (clone L161), anti-CD8 (clone SK1), anti-CD14 (clone M5E2), anti-CD11c (clone 3.9), anti-CD4 (clone OKT4), anti-CD20 (clone 2H7), anti-CD123 (clone 7G3), anti-HLA-DR (clone Immu-357), anti-CD3 (clone SP34–2), anti-CCR6 (clone GO34E3), anti-PD1 (clone EH12.2H7), anti-CD28 (clone 28.2), anti-CD69 (clone FN50), anti-CD95 (clone DX2), anti-CDCXCR3 (clone G025H7), anti-CD25 (clone M-A251), anti-Ki67 (clone B56), anti-CXCR5 (clone 1C6), and anti-CCR7 (clone 3D12). A minimum of 100,000 CD3+ T cells were collected, and post-acquisition analysis performed by Y.C. using FlowJo (Version 9.6, TreeStar) software.
CAI Y., Watkins M.A., Xue F., Ai Y., Cheng H., Midkiff C.C., Wang X., Alvarez X., Poli A.N., Salvino J.M., Veazey R.S, & Montaner L.J. (2019). BCL6 BTB-specific Inhibition via FX1 Treatment Reduces Tfh Cells & Reverses Lymphoid Follicle Hyperplasia in Indian Rhesus Macaque (Macaca mulatta). Journal of medical primatology, 49(1), 26-33.
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2

Comprehensive B Cell Immunophenotyping

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For analysis of B cells from blood, cultures or nasal tissue, cell samples were simultaneously Fc-blocked with unlabeled mouse IgG (Lampire) and ICAM-1-blocked with anti-ICAM-1, (BioLegend), which was fluorescently-labeled or not, depending upon the experiment. After 30 minutes at 4°C, B cells were stained with fluorescently tagged virus (Alexa Fluor 488-RV-A39 and Alexa Fluor 568-RV-A16), viability dye Live/Dead Aqua (ThermoFisher), and various combinations of the following fluorescent antibodies depending on the sample type and application: anti-CD3 (BioLegend), anti-CD11c (BioLegend), anti-CD19 (BioLegend), anti-CD20 (BioLegend), anti-CD27 (ThermoFisher), anti-CD38 (Becton Dickinson), anti-CCR5 (ThermoFisher), anti-CXCR3 (BioLegend), anti-CXCR5 (BioLegend), anti-IgD (ThermoFisher), anti-IgM (BioLegend), anti-IgG (BD Biosciences), anti-IgA (Miltenyi), and anti-IgE (BioLegend). After incubating for 30 minutes at 4°C, cells were then fixed and permeabilized (FoxP3 fix/perm kit, ThermoFisher), before staining for intracellular IgM (BioLegend), IgG (Becton Dickinson), IgA (Miltenyi), IgE (BioLegend), Ki-67 (BioLegend), and T-bet (BioLegend). Cells were analyzed on an LSR Fortessa Cytometer (Becton Dickinson) using FlowJo version 10.5.3 (TreeStar). (See Table S1).
Eccles J.D., Turner R.B., Kirk N.A., Muehling L.M., Borish L., Steinke J.W., Payne S.C., Wright P.W., Thacker D., Lahtinen S.J., Lehtinen M.J., Heymann P.W, & Woodfolk J.A. (2020). T-bet+ Memory B Cells Link to Local Cross-Reactive IgG upon Human Rhinovirus Infection. Cell reports, 30(2), 351-366.e7.
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3

Comprehensive Antibody Validation for Immunoblotting and Flow Cytometry

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Anti-mouse β-actin (AF0003; 1:1,000 dilution), Horseradish peroxidase (HRP)-anti-rabbit immunoglobulin G (IgG) (A0208; 1:3,000 dilution) and HRP-anti-mouse IgG (A0216; 1:3,000 dilution) were purchased from Beyotime Institute of Biotechnology (Haimen, China). Anti-ACK1 (PA5-102625) antibody was purchased from Invitrogen (Carlsbad, CA, USA). Anti-p-p65 (9246), anti-p65 (9242), anti-p-Erk (9101), anti-Erk (9107), anti-p-JNK (9251), anti-JNK (9251), anti-p-p38 (4511) and anti-p38 (9228) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). For flow cytometry purposes, fluorochrome-conjugated anti-B220 (Cat#: 103206), anti-CD4 (Cat#: 100406), anti-CD40 (Cat#: 124610), anti-F4/80 (Cat#: 123108), anti-CD86 (Cat#: 105012), anti-CD11c (Cat#: 117310), anti-GL7 (Cat#: 144608), anti-CD95 (Cat#: 152604), anti-CXCR5 (Cat#:145506), and anti-PD-1 (Cat#:135206) antibodies were purchased from BioLegend. Unless specifically listed otherwise, all flow cytometry antibodies were stained at a 1:100 dilution.
Jing L., Zhang X., Liu D., Yang Y., Xiong H, & Dong G. (2022). ACK1 Contributes to the Pathogenesis of Inflammation and Autoimmunity by Promoting the Activation of TLR Signaling Pathways. Frontiers in Immunology, 13, 864995.
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4

Comprehensive Immune Cell Profiling

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The cells were resuspended in ice-cold PBS containing 1% FBS and were stained with anti-CD3, anti-CD4, anti-CD8, anti-B220, anti-CD38, anti-CD138, anti-PD-1, anti-CXCR5, anti-CD62L, anti-OX40, anti-IgD, anti-CD95, anti-GL7, anti-CD11c, anti-CD80, anti-CD86, anti-MHC II, anti-CD40, anti-CD69, anti-CD11b, anti-Gr-1, and anti-F4/80 antibodies (Biolegend). Intracellular staining of IFN-γ, TNF-α, IL-17a, Helios, and Foxp3 was performed after 4 h of stimulation with PMA and ionomycin following a standard protocol (BD Biosciences). The results were obtained on a BD FACS Fortessa flow cytometer and were analyzed using FlowJo software.
Xiao Z.X., Hu X., Zhang X., Chen Z., Wang J., Jin K., Cao F.L., Sun B., Bellanti J.A., Olsen N, & Zheng S.G. (2020). High salt diet accelerates the progression of murine lupus through dendritic cells via the p38 MAPK and STAT1 signaling pathways. Signal Transduction and Targeted Therapy, 5, 34.
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5

Comprehensive B Cell Immunophenotyping

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For analysis of B cells from blood, cultures or nasal tissue, cell samples were simultaneously Fc-blocked with unlabeled mouse IgG (Lampire) and ICAM-1-blocked with anti-ICAM-1, (BioLegend), which was fluorescently-labeled or not, depending upon the experiment. After 30 minutes at 4°C, B cells were stained with fluorescently tagged virus (Alexa Fluor 488-RV-A39 and Alexa Fluor 568-RV-A16), viability dye Live/Dead Aqua (ThermoFisher), and various combinations of the following fluorescent antibodies depending on the sample type and application: anti-CD3 (BioLegend), anti-CD11c (BioLegend), anti-CD19 (BioLegend), anti-CD20 (BioLegend), anti-CD27 (ThermoFisher), anti-CD38 (Becton Dickinson), anti-CCR5 (ThermoFisher), anti-CXCR3 (BioLegend), anti-CXCR5 (BioLegend), anti-IgD (ThermoFisher), anti-IgM (BioLegend), anti-IgG (BD Biosciences), anti-IgA (Miltenyi), and anti-IgE (BioLegend). After incubating for 30 minutes at 4°C, cells were then fixed and permeabilized (FoxP3 fix/perm kit, ThermoFisher), before staining for intracellular IgM (BioLegend), IgG (Becton Dickinson), IgA (Miltenyi), IgE (BioLegend), Ki-67 (BioLegend), and T-bet (BioLegend). Cells were analyzed on an LSR Fortessa Cytometer (Becton Dickinson) using FlowJo version 10.5.3 (TreeStar). (See Table S1).
Eccles J.D., Turner R.B., Kirk N.A., Muehling L.M., Borish L., Steinke J.W., Payne S.C., Wright P.W., Thacker D., Lahtinen S.J., Lehtinen M.J., Heymann P.W, & Woodfolk J.A. (2020). T-bet+ Memory B Cells Link to Local Cross-Reactive IgG upon Human Rhinovirus Infection. Cell reports, 30(2), 351-366.e7.
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