THP1 cells with or without knockdown were labelled with 5 µg/ml Calcein AM (C3100MP, Molecular Probes, Eugene, OR, USA) and incubated on top of a monolayer of HUVECs for 30 min at 37°C. Non‐adhering cells were washed away by multiple washing steps with PBS after which the cells were lysed in Triton‐X 0.5% for 10 min. Fluorescence was measured at λex 485 nm and λem 514 nm. Each condition was performed in triplicate. In case of cell stimulation, THP1 cells were stimulated with 500 ng/ml recombinant EphrinB1 (7654‐EB, R&D Systems, Minneapolis, MN, USA) or EphrinB2 (7397‐EB, R&D Systems, Minneapolis, MN, USA) for 30 min before addition to the monolayer of endothelial cells.
Calcein am c3100mp
Manufactured by Thermo Fisher Scientific
Sourced in United States
Calcein AM (C3100MP) is a fluorescent dye used for the detection and quantification of live cells. It is a non-fluorescent, cell-permeant compound that is converted to a bright green-fluorescent calcein upon hydrolysis by intracellular esterases, indicating the presence of living cells.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using calcein am c3100mp
1
Leukocyte-Endothelial Adhesion Assay
2
Monocyte Adhesion to Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
THP-1 monocytes were labelled with 5 μM calcein-AM (C3100MP; Molecular Probes, Oregon, USA) for 30 min and then seeded (2.5 × 105 cells) over hECs previously exposed to Tyr or its metabolites (100 μM) for 16 h and to TNF-α (10 ng mL -1 ) for additional 6 h. After the co-culture, the cells were washed with PBS and fluorescence was measured at excitation and emission wavelengths of 485 nm and 530 nm, respectively, using a Fluoroskan Microplate Fluorometer (ThermoFisher Scientific). THP-1 cells adhered to hECs were visualized by fluorescence microscopy with a motorized inverted fluorescence microscope IX81 equipped with a FITC filter and a Megaview-II digital camera (Olympus).
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!