Pe conjugated anti cd62l

Manufactured by BioLegend

PE-conjugated anti-CD62L is a monoclonal antibody that recognizes the CD62L cell surface antigen. CD62L, also known as L-selectin, is a cell adhesion molecule involved in lymphocyte homing and migration. The PE (phycoerythrin) fluorescent label allows for the detection and quantification of CD62L-expressing cells using flow cytometry.

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3 protocols using pe conjugated anti cd62l

Multiparameter Flow Cytometry Analysis of Lymphocyte Subsets

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Cells were harvested and stimulated for 4 h with PMA (50 ng/ml), ionomycin (750 ng/ml), and Brefeldin A (BioLegend). For intracellular staining, cells were fixed and permeabilized (eBioscience) and then stained with FITC-conjugated anti–IFN-γ (505806, BioLegend), PE-conjugated anti–IL-13 (12-7133-81, eBioscience), PerCP/Cy5.5-conjugated anti–IL-17A (506919, BioLegend), and APC-conjugated anti-FOXP3 (17-5773-80, eBioscience) antibodies. To analyze lymphocyte development in Pten^fl/flIl17a^cre mice, cells isolated from the thymus, pLNs, and spleen were stained with FITC-conjugated anti-CD3ε (11-0031-81, eBioscience), PE-conjugated anti-B220 (12-0451-82, eBioscience), PerCP/Cy5.5-conjugated anti-CD8 (126609, BioLegend), APC-conjugated anti-CD4 (100411, BioLegend), FITC-conjugated anti-CD44 (103006, BioLegend), PE-conjugated anti-CD62L (104407, BioLegend), and PerCP/Cy5.5-conjugated anti-TCR γ/δ (118117, BioLegend) antibodies. For other surface antigen staining, cells were stained with APC-conjugated anti–IL-6Rα (115811, BioLegend), APC-conjugated anti-CD45 (103111, BioLegend), FITC-conjugated anti–GR-1 (108405, BioLegend), and FITC-conjugated anti-CD11b (101205, BioLegend) antibodies. Stained cells were then analyzed using a FACSCalibur flow cytometer (BD Bioscience), and data were analyzed with FlowJo software.

Kim H.S., Jang S.W., Lee W., Kim K., Sohn H., Hwang S.S, & Lee G.R. (2017). PTEN drives Th17 cell differentiation by preventing IL-2 production. The Journal of Experimental Medicine, 214(11), 3381-3398.

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Flow Cytometry Analysis of Murine Lymphocytes

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Conjunctivae tissues were first digested in RPMI (Invitrogen) with 2mg/ml DNase and 2mg/ml Collagenase (Roche) at 37°C. The following Abs were used for flow cytometry analysis: FITC-conjugated anti-CD4, PerCP-Cy5.5-conjugated anti-CD44, PE-conjugated anti- IL-7Rα, APC-conjugated anti- IL-15Rα, PE- conjugated anti-CD62L, FITC-conjugated anti-Ki-67, Brilliant Violet 421-conjugated anti-Annexin V (BioLegend), PE-Cy7-conjugated anti- IL-17, and PE-conjugated anti- IL-17 (eBioscience). For intracellular IL-17 staining, cells were stimulated with 50 ng/mL phorbol 12-myristate 13-acetate and 500 ng/mL ionomycin (Sigma-Aldrich) for 6 hours at 37°C and 5% CO₂ in the presence of GolgiStop™ (4 μl per 6 mL cell culture, BD Biosciences) to inhibit cytokine secretion. Stained cell were examined with an LSR II flow cytometer (BD Biosciences), and the results were analyzed using FlowJo software (Tree Star).

Chen Y., Chauhan S.K., Tan X, & Dana R. (2016). Interleukin-7 and -15 Maintain Pathogenic Memory Th17 Cells in Autoimmunity. Journal of autoimmunity, 77, 96-103.

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Neutrophil CD125 and CD62L Regulation

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Upregulation of surface CD125 was assessed after stimulating neutrophils for 30 min with either IL-5 (30 ng/mL), N-formyl methionine-leucyl-phenylalanine (fMLP; 10 μM) with cytochalasin b (1 μg/mL), or phorbol myristate acetate (PMA; 50 ng/mL) and ionomycin (1 μg/ml) (Sigma Aldrich, St. Louis, MO). Downregulation of surface CD62L expression after stimulation with IL-5 (10-100 ng/mL) for 15 min was assessed via flow cytometry using PE-conjugated anti-CD62L (BioLegend).

Lawrence M.G., Teague W.G., Feng X., Welch C., Etter E., Negri J., Spano M., Wavell K., Braciale T., Steinke J.W, & Borish L. (2021). Interleukin-5 Receptor alpha (CD125) Expression on Human Blood and Lung Neutrophils. Annals of allergy, asthma & immunology : official publication of the American College of Allergy, Asthma, & Immunology, 128(1), 53-60.e3.

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