Starting in mid July, we are going to provide 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 human cytokines:
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- Thaw samples on ice. Centrifuge @ 10000 g for 10 min at 4oC.
- 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 adding 100 ul of wash buffer. Vaccum.
- Add 50 ul of diluted sample & 25 ul of mixed beads to each well. Seal plate and shake at 500 rpm for 2 hours RT.
- Wash with 200 ul of wash buffer and vaccum.
- 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 200 ul of wash buffer and vaccum.
- 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 $40. We will be running 39 samples plus 8 standard curve titrations in duplicate on a 96 well plate.
To order this service, please complete an order for Service_Cytokine Assay under the Flow Cytometry tab on PPMS:
https://au.ppms.info/wrh/order/?pf=2
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