C. trachomatis LGV-L2 RifR pGFP::SW2, WT C. trachomatis LGV-L2, RST5 LGV-L2, RST17 LGV-L2, or Targetron-mediated CPAF mutant LGV-L2 was grown and purified as previously described49 (link),51 (link),63 (link). Each preparation was titered uniformly by counting inclusion forming units on monolayers of Vero cells. C. trachomatis was diluted in RPMI with 10% FBS and added at MOI 5 in 100 µl in 96-well plates, mixed via pipetting, and centrifuged onto cells at 1500×g for 30 min at 4 °C. At 27, 46, and 70 h, cells were mixed and 25 µl was used for flow cytometry quantification of C. trachomatis burden and cell death. At 70 h, 25 µl of cell culture supernatant was measured by custom Luminex assay for 17 human cytokines (Millipore), or 50 µl of cell culture supernatant was measured by ELISA for [CXCL10] or [RANTES] (R&D).
Luminex assay for 17 human cytokines
Manufactured by Merck Group
The Luminex assay for 17 human cytokines is a multiplex platform that allows for the simultaneous detection and quantification of 17 different cytokines from a single sample. The assay utilizes color-coded magnetic beads coated with specific antibodies to capture and measure the target cytokines. This technology enables efficient and reliable analysis of complex biological samples.
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Lab products found in correlation
2 protocols using luminex assay for 17 human cytokines
1
Quantification of Chlamydia Infection and Immune Response
2
Infection and Cytokine Analysis of C. trachomatis
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C. trachomatis L2-GFP49 (link) was a gift from the Derre Lab and the Valdivia lab. Elementary bodies (EBs) were purified on Omnipaque-350 gradients as previously published.50 (link) Each preparation was titered by counting inclusion formation units by microscopy on monolayers of Vero cells. C. trachomatis was diluted in RPMI with 10% FBS and added at either MOI 0.5 for HeLa cells or MOI 5 for LCLs in 100 μL in 96-well plates as indicated in text. Cells were mixed via pipetting and centrifuged onto cells at 1,500 × g for 30 min at 4°C. Infected cells were then incubated at 37°C. For suspension cells, at 24, 48, and 72 h, cells were mixed and 15 μL of cells were used for flow cytometry quantification of C. trachomatis infection levels. At 72 h, supernatant was collected to measure cytokine production by either Luminex assay for 17 human cytokines (Millipore) or enzyme-linked immunosorbent assay (ELISA) for [IFNα2] or [IL-6] (R&D).
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