Anti ccl20

Manufactured by R&D Systems

Anti-CCL20 is a recombinant protein that binds to the chemokine CCL20. CCL20 is involved in the recruitment of T cells, dendritic cells, and B cells to sites of inflammation. Anti-CCL20 can be used in research applications to study the role of CCL20 in various biological processes.

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Lab products found in correlation

Cytofix cytoperm plus kit, by BD (3 mentions) Lsrii system, by BD (2 mentions) Anti rorγt, by Thermo Fisher Scientific (2 mentions) Golgiplus, by BD (2 mentions) Anti ifn γ, by R&D Systems (2 mentions) Anti il 17f, by R&D Systems (2 mentions) Anti il 17a, by Thermo Fisher Scientific (2 mentions) Anti t bet, by Thermo Fisher Scientific (2 mentions) Anti il 22, by R&D Systems (2 mentions) Fortessa flow cytometer, by BD (2 mentions) Foxp3 staining buffer set, by Thermo Fisher Scientific (2 mentions) Facs caliber, by BD (2 mentions) Anti il 6, by R&D Systems (2 mentions) Anti il 1β, by BioXCell (1 mentions) Anti il 1α, by BioXCell (1 mentions) Gentlemacs, by Miltenyi Biotec (1 mentions) Anti tnf α, by BioLegend (1 mentions) Anti il 17a, by BioLegend (1 mentions) Anti gm csf, by BioLegend (1 mentions) Fortessa, by BD (1 mentions)

5 protocols using anti ccl20

Candida Albicans Oral Infection Model

Cited 4 times

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OPC was induced by sublingual inoculation with a cotton ball saturated in C. albicans for 75 min, as described (5 (link)). For re-challenge, mice were infected 6 weeks after the primary infection (8 (link)). Tongue homogenates were prepared on a GentleMACS (Miltenyi Biotec, Auburn, CA) and CFU determined by serial dilutions on YPD agar. Anti-CCL20 (R&D Systems, Minneapolis, MN), anti-IL-1α, anti-IL-1β or isotype control Abs (BioXCell, West Lebanon, NH) were administered on day -1 p.i. (100–1000 μg/mouse). CAF2-1 or Bwp17 C. albicans strains (derived from SC5314, www.candidagenome.org/Strains.shtml) were used as wild-type. Knockout strains (sap4-6Δ/Δ ece1Δ/Δ, efg1Δ/Δ) and HUN96 were described (16 (link), 29 (link), 34 (link), 35 (link)).

Verma A.H., Richardson J.P., Zhou C., Coleman B.M., Moyes D.L., Ho J., Huppler A.R., Ramani K., McGeachy M.J., Mufazalov I.A., Waisman A., Kane L.P., Biswas P.S., Hube B., Naglik J.R, & Gaffen S.L. (2017). Oral epithelial cells orchestrate innate Type 17 responses to Candida albicans through the virulence factor Candidalysin. Science immunology, 2(17), eaam8834.

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Stimulation and Cytokine Profiling of T Cells

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T cells purified by cell sorting were stimulated with Leukocyte Activation Cocktail, with GolgiPlus^™ (BD Pharmingen) for 6 h at 37°C under 5% CO₂ before being stained with following antibodies alone or in combinations: anti-IL-17A (eBioscience), anti-IL-17F, anti-IL-22, anti-IL-6, anti-CCL20 and anti-IFNγ (R&D Systems) by using Cytofix/CytoPerm Plus kit (BD Pharmingen) or anti-RORγt, anti-FOXP3, anti-T-bet or anti-GATA3 using Foxp3 staining buffer set (eBioscience) according to the manufacturer’s instructions. All flow cytometry was done using FACSCaliber, LSR II System or Fortessa flow cytometers (BD Biosciences), and the data were subsequently analyzed using FlowJO software (TreeStar).

Singh S.P., Parween F., Edara N., Zhang H.H., Chen J., Otaizo-Carrasquero F., Cheng D., Oppenheim N.A., Ransier A., Zhu W., Shamsaddini A., Gardina P.J., Darko S.W., Singh T.P., Douek D.C., Myers T.G, & Farber J.M. (2023). Human CCR6+ Th cells show both an extended stable gradient of Th17 activity and imprinted plasticity. Journal of immunology (Baltimore, Md. : 1950), 210(11), 1700-1716.

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Comprehensive Cytokine Analysis of Activated T Cells

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T cells purified by cell sorting were stimulated with Leukocyte Activation Cocktail, with GolgiPlus (550583; BD Pharmingen) for 6 h at 37°C with 5% CO₂ before being stained with following antibodies alone or in combinations: anti-IL-17A (512305), anti-IFN-γ (502530), BioLegend), anti-TNF-α (502944), anti-GM-CSF (502305) all from BioLegend and anti-CCL20 (C360A; R&D Systems) by using Cytofix/CytoPerm Plus kit (555028; BD Pharmingen). Samples were analyzed using a Fortessa flow cytometer (BD Biosciences), and the data were analyzed using FlowJo software (BD Biosciences).

Parween F., Singh S.P., Zhang H.H., Kathuria N., Otaizo-Carrasquero F.A., Shamsaddini A., Gardina P.J., Ganesan S., Kabat J., Lorenzi H.A., Myers T.G, & Farber J.M. (2023). Chemokine positioning determines mutually exclusive roles for their receptors in extravasation of pathogenic human T cells. bioRxiv.

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Transwell assay for Treg migration

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Isolated naïve T cells were seeded in six‐well plates with anti‐CD3 (0.5 μg/m:, ebioscience) and anti‐CD28 (2 μg/mL, ebioscience) stimulated for 2 days. Then, cocultured with NPC cells with transwell in the presence or absence of anti‐TGF‐β1 (10 ng/ml, R&D System) for 4 days.
Migration of Tregs was tested by 5 μm pore‐size transwell (Corning Costar). 1 × 10⁴ Tregs were seeded in upper chamber with NPC cells supernatant in lower chamber and cultured with or without anti‐CCL20 (10 ng/mL, R&D System) for 24 hours. The number of migrated cells in the lower chamber was counted by microscope.

Wang J., Luo Y., Bi P., Lu J., Wang F., Liu X., Zhang B, & Li X. (2020). Mechanisms of Epstein‐Barr virus nuclear antigen 1 favor Tregs accumulation in nasopharyngeal carcinoma. Cancer Medicine, 9(15), 5598-5608.

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Intracellular Cytokine Analysis of T Cells

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T cells purified by cell sorting were stimulated with Leukocyte Activation Cocktail, with GolgiPlus ™ (BD Pharmingen) for 6 h at 37°C under 5% CO 2 before being stained with following antibodies alone or in combinations: anti-IL-17A (eBioscience), anti-IL-17F, anti-IL-22, anti-IL-6, anti-CCL20 and anti-IFNγ (R&D Systems) by using Cytofix/CytoPerm Plus kit (BD Pharmingen) or anti-RORγt, anti-FOXP3, anti-T-bet or anti-GATA3 using Foxp3 staining buffer set (eBioscience) according to the manufacturer's instructions. All flow cytometry was done using FACSCaliber, LSR II System or Fortessa flow cytometers (BD Biosciences), and the data were subsequently analyzed using FlowJO software (TreeStar).

Singh S.P., Parween F., Edara N., Zhang H.H., Chen J., Otaizo-Carrasquero F., Cheng D., Oppenheim N.A., Ransier A., Zhu W., Shamsaddini A., Gardina P.J., Darko S.W., Singh T.P., Douek D.C., Myers T.G, & Farber J.M. (2023). Human CCR6+ Th Cells Show Both an Extended Stable Gradient of Th17 Activity and Imprinted Plasticity. Journal of immunology (Baltimore, Md. : 1950), 210(11).

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