Af503

Manufactured by R&D Systems

The AF503 is a laboratory instrument designed for the detection and quantification of biological analytes. It utilizes a specialized detection method to provide accurate measurements. The core function of the AF503 is to facilitate precise analysis of samples in a research or diagnostic setting.

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Lab products found in correlation

Af478, by R&D Systems (2 mentions) Protein g sepharose beads, by Abcam (2 mentions) Immun star westernc kit, by Bio-Rad (1 mentions) Quantity one software, by Bio-Rad (1 mentions)

3 protocols using af503

Depletion of Chemokines from Conditioned Medium

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MEPs or Lin- cells depleted of MEPs (1 × 10⁶ cells/ml) were cultured in DMEM serum-free medium for 24 hours to generate conditioned medium (CM). CM (1 ml) was depleted of CCL5 or CXCL16 by overnight incubation at 4 °C with 1 µg anti-CCL5 (AF478, R&D Systems) or anti-CXCL16 (AF503, R&D Systems) antibodies, respectively. The CM was then incubated with 50 µl Protein G Sepharose® beads (ab193259, Abcam) for 1 h at 4 °C. Beads were removed by centrifugation in 2000 g for 10 min.

Vorontsova A., Cooper T.J., Haj-Shomaly J., Benguigui M., Levin S., Manobla B., Menachem R., Timaner M., Raviv Z, & Shaked Y. (2023). Chemotherapy-induced tumor immunogenicity is mediated in part by megakaryocyte-erythroid progenitors. Oncogene, 42(10), 771-781.

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Depletion of Cytokines from Cell-Conditioned Medium

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MEPs or Lin-cells depleted of MEPs (1x10⁶ cells/ml) were cultured in DMEM serum-free medium for 24 hours to generate conditioned medium (CM). CM (1 ml) was depleted of CCL5 or CXCL16 by overnight incubation at 4°C with 1μg anti-CCL5 (AF478, R&D Systems) or anti-CXCL16 (AF503, R&D Systems) antibodies, respectively. The CM was then incubated with 50 μl Protein G Sepharose® beads (ab193259, Abcam) for 1 hour at 4°C. Beads were removed by centrifugation in 2000g for 10 min.

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Quantitative Western Blot Analysis of CXCL16

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Protein samples were separated on 10% SDS-polyacrylamide gel and analyzed by Western immunoblot using a mouse CXCL16 antibody (0.2 μg/ml; R&D System, AF503) and HRP-tagged rabbit anti goat IgG secondary antibody (1:2000; Dako), and subsequently detected using a commercial chemiluminescent assay (Immun-Star WesternC Kit; Bio-Rad). Densitometric analysis was performed with Quantity One software (Biorad). Interpretation of western-blot bands for CXCL16 was according to Gough et al. (2004 (link)).

Rosito M., Lauro C., Chece G., Porzia A., Monaco L., Mainiero F., Catalano M., Limatola C, & Trettel F. (2014). Trasmembrane chemokines CX3CL1 and CXCL16 drive interplay between neurons, microglia and astrocytes to counteract pMCAO and excitotoxic neuronal death. Frontiers in Cellular Neuroscience, 8, 193.

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