The plasma concentration of 27 cytokines, growth factors or soluble receptors was determined by enzyme-linked immunosorbent assays (ELISA) using two Fluorokine MAP kits (the human angiogenesis panel A and the human high sensitivity cytokine panel; R&D Systems, Abingdon, UK). Competition experiments were conducted to test interference of aflibercept with the detection of VEGFA, VEGFD and placental growth factor (PlGF). Angiopoietin-2 (ANGPT2), SDF1-α, HGF, VEGFC, soluble VEGF receptor 3 (sVEGFR3) and sVEGFR2 were assessed by single ELISA (R&D Systems). Plasma markers were analysed at baseline, at 30 and 60 days after the first study treatment infusion and 30 days after the last aflibercept infusion.
Fluorokine map kit
Manufactured by R&D Systems
Sourced in United Kingdom
The Fluorokine MAP kit is a multiplex immunoassay system designed to detect and quantify multiple analytes in a single sample. It utilizes fluorescently-labeled magnetic beads to measure the concentrations of target proteins simultaneously.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using fluorokine map kit
1
Plasma Cytokine Profiling in Aflibercept Study
2
Inflammatory Cytokine Response Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples collected during the clinical intervention (baseline, 1, 2, 4, 6 and 8 h) into heparin tubes were immediately cultured as follows: 500-μl blood aliquots were mixed with 500 μl of Roswell Park Memorial Institute (RPMI) medium containing antibiotics on a twenty-four-well plate, and lipopolysaccharide (LPS) (1 μg/ml) or vehicle (control group) was added before incubation at 37°C for 24 h. At the end of the culture period, samples were centrifuged at 2000 g for 5 min, and the supernatants were collected and kept at -20°C until analysis for inflammatory cytokine levels. The supernatants were collected and stored at -20°C. Cytokines (IL-1β, IL-6, IL-8, IL-10 and TNF-α) in the supernatants were measured with Luminex xMAP Technology using commercially available Fluorokine MAP kits (R&D systems), and data were analysed on the xPONENT software. Final data are presented as the difference between LPS-treated and unstimulated control.
3
Quantifying MMP and TIMP Levels by ELISA and Luminex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MMP and TIMP concentrations were analyzed by ELISA (Duoset; R&D Systems, Abingdon, U.K.) or by Luminex bead array (Luminex 200; Bio-Rad, Hertfordshire, U.K.) using the Fluorokine MAP kit (R&D Systems) according to the manufacturer’s instructions. Lower limits of sensitivity for the Duoset kits are: 21.2 pg/ml for TIMP-1, 31.2 pg/ml for TIMP-2, 156 pg/ml for MMP-1, and 31.2 pg/ml for MMP-10. In the Fluorokine Luminex kit the lower limits are: 1.1 pg/ml for MMP-1, 12.6 pg/ml for MMP-2, 7.3 pg/ml for MMP-3, 6.6 pg/ml for MMP-7, 13.7 pg/ml for MMP-9, and 3.2 pg/ml for MMP-10. Variation on secreted concentrations of MMPs and TIMP between individual donors is shown in Supplemental datasets .
4
ELISA and Luminex Assays for MMP and TIMP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At 24 h post-stimulation, MMP and TIMP concentrations were analyzed by ELISA (Duoset, R&D Systems, Abdingdon, UK) or by Luminex bead array (Luminex 200, Bio-Rad, Hertfordshire, UK) using the Fluorokine MAP kit (R&D Systems) according to manufacturer’s instructions. Lower limits of sensitivity for the Duoset kits are: 21.2 pg/ml for TIMP-1, 31.2 pg/ml for TIMP-2, and 156 pg/ml for MMP-1. In the Fluorokine Luminex kit, the lower limits are: 1.1 pg/ml for MMP-1, 12.6 pg/ml for MMP-2, 7.3 pg/ml for MMP-3, and 13.7 pg/ml for MMP-9. Samples were within the linear range of detection as indicated by the manufacturer.
5
ELISA and Luminex Quantification of MMPs and TIMPs
6
Multiplex Immunoassay for MMPs and Adhesion Molecules
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Microparticle based multiplex immunoassay were used to detect MMPs and soluble adhesion molecules and were analyzed on the Luminex 200 (BioRad, Hertfordshire, UK). MMPs were analysed using the Fluorokine MAP kit (R&D Systems) according with the manufacturer’s instructions. Lower limits of sensitivity are: 1.1 pg/ml for MMP-1, 12.6 pg/ml for MMP-2, 7.3 pg/ml for MMP-3, 6.6 pg/ml for MMP-7, 16.6 pg/ml for MMP-8, 13.7 pg/ml for MMP-9, and 3.2 pg/ml for MMP-10. All samples were run with appropriate controls and were within the linear range of detection as indicated by the manufacturer. Adhesion molecules were analyzed using the 4-plex human adhesion molecule performance kit (R&D Systems). Lower limits of sensitivity are: 303 pg/ml for ICAM-1, 7.4 pg/ml for E-Selectin, 12.2 pg/ml for P-Selectin and 529 pg/ml for ICAM-1.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!