Starting in mid July , 2020 we are going to provide providing the cytokine multiplex assay as a new service. This assay is validated for serum, plasma and culture supernatant.
With as little as 12.5 µl serum/plasma, or 25 µl for tissue culture samples per well, this panel allows simultaneous quantification of 13 12 human cytokines:
Details of the kit can be found here:
|https://www.biolegend.com/en-us/products/legendplex-humanhu-th-cytokine-panel-1312-plex-filterwfp-platev02-1579119471
Brief protocol:
- Thaw samples on ice. Centrifuge @ 12000g for 5min.
- Dilute 50 ul of serum samples 1:1 with assay buffer. Cell culture supernatant samples can be tested without dilutions but may need a preliminary experiment to determine the appropriate dilution factor.
- Prepare 1:4 serial dilution of standards.
- Pre-wet filter plate by dispensing 100 ul of wash buffer using the plate washer. Vacuum.
- Add 25 ul of assay buffer to each well. When pipetting aim for the side of the wells and avoid touching the membrane. This can help preventing leakages.
- Add 50 ul of diluted sample/standards & 25 ul of mixed beads to each well.
- Seal plate and shake at 500 rpm for 2 hours RT.
- Wash with dispensing 200 ul of wash buffer using the plate washer and vacuum.
- Add 25 ul of detection antibodies to each well. Seal plate and shake at 500 rpm for 1 hour RT.
- Add 25 ul of SA-PE to each well. Seal plate and shake at 500 rpm for 30 min RT.
- Wash with dispensing 200 ul of wash buffer using the plate washer and vacuum.
- Add 150 ul of wash buffer to each well. Resuspend beads on a plate shaker for 1 min.
- Samples are now ready to read on a cytometer.
Cost:
Each sample will cost ~$58 (subject to updates). We will be running 40 samples (including 1 assay control) plus 8 standard curve titrations in duplicate on a 96 well plate.
We'll also include a frozen aliquot of sample standard to monitor variation between plates.
To order this service, please complete an order for Service_Cytokine Assay under the Flow Cytometry tab on PPMS:
...