PMA/ionomycin-stimulated PBMCs were stained using fixable AQUA live/dead stain (Invitrogen) and the following antibodies: CD3-BUV395 (Invitrogen), CD4-BV605 (BD Biosciences), CD45RA-PerCPVio700 (BD Biosciences), CD31-BV421 (BD Biosciences), CD45RO-PE (BD Biosciences), and CD8-APC-Cy7 (BD Biosciences) in FACS buffer (phosphate-buffered saline containing 0.2% bovine serum albumin and 0.02% sodium azide). PBMCs were fixed and permeabilized (Foxp3, eBioscience). The PBMCs were intracellular stained with Ki67-AF647 (BD Pharmigen, measured in un-stimulated PBMCs) and IL-8-AF488 (CXCL8, BD Biosciences, measured in stimulated PBMCs) and incubated in the dark on ice for 1 hour. Samples were analyzed on an LSRII (Becton Dickinson) using FACSDiva software v.8.0. Subsequent data analysis was performed using FlowJo software 10.4.
Il 8 af488
Manufactured by BD
The IL-8-AF488 is a fluorescently labeled recombinant human interleukin-8 (IL-8) protein. It is a chemokine that plays a role in the immune response and inflammation. The AF488 dye is covalently attached to the IL-8 protein, allowing for its detection and quantification using fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Cd8 apc cy7, by BD (2 mentions)
Cd4 bv605, by BD (2 mentions)
Aqua live dead stain, by Thermo Fisher Scientific (2 mentions)
Cd45ro pe, by BD (2 mentions)
Cd31 bv421, by BD (2 mentions)
Cd3 buv395, by Thermo Fisher Scientific (2 mentions)
Cd45ra percpvio700, by BD (2 mentions)
Ki67 af647, by BD (2 mentions)
2 protocols using il 8 af488
1
PBMC immunophenotyping and activation
2
Multiparametric Flow Cytometry Analysis of PBMCs
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PMA/ionomycin-stimulated PBMCs were stained using fixable AQUA live/dead stain (Invitrogen) and the following antibodies: CD3-BUV395 (Invitrogen), CD4-BV605 (BD Biosciences), CD45RA-PerCPVio700 (BD Biosciences), CD31-BV421 (BD Biosciences), CD45RO-PE (BD Biosciences), and CD8-APC-Cy7 (BD Biosciences) in FACS buffer (phosphate-buffered saline containing 0.2% bovine serum albumin and 0.02% sodium azide). PBMCs were fixed and permeabilised (Foxp3, eBioscience). The PBMCs were intracellular stained with Ki67-AF647 (BD Pharmigen, measured in un-stimulated PBMCs) and IL-8-AF488 (CXCL8, BD Biosciences, measured in stimulated PBMCs) and incubated in the dark on ice for 1 hour. Samples were analyzed on an LSRII (Becton Dickinson) using FACSDiva software v.8.0. Subsequent data analysis was performed using FlowJo software 10.4. For gating strategy see Figure 1SB
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