PPT Slide

Simultaneous quantitation of secreted IL-8 and cell surface ICAM-1after stimulation with TNF-a

• Plate 5,000 HUVEC cells/well in 96 well plate

• Incubate overnight

• Replace media with 100 ?l fresh media with or without 10 ng/ml TNF-a and incubate overnight

• Add 50 ml PBS with NaN3 containing:

beads coated with capture anti-IL-8 antibody

dyeA-labeled detection anti-IL-8 antibody

dyeB-labeled anti-ICAM-1 antibody

• Incubate overnight at room temperature

• Scan

FLISA with Cells in the Well

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