Product: Brilliant Violet 421™ anti-mouse/human IL-5

Catalog No.: 504311

Hidde Douna, MSc LACDR

Brilliant Violet 421™ anti-mouse/human IL-5
Overall:	Product Quality:	Ease of Use:
Difficult signal to pick up.

Experimental Design

Application:	Flow cytometry
Cells used:	Mouse splenocytes
Brief Protocol:	Splenocytes were stimulated with PMA/ionomycin/Brefeldin A for 5h. Cells were then fixed/permeabilized and 2e5 splenocytes were stained in a final volume of 25 microliter using a 1:100 dilution of the BV421 IL-5 antibody. Incubation period was 30 minutes at 4 degrees.
Results Summary:	No signal found.
Additional Notes:	While IFNy, IL-17 and IL-10 could be clearly detected, absolutely no signal for IL-5 was detected.

Data Image Click on the image for full size data.

Publication Status This data has not been published.

Technical Service Notes

Based on our QC testing, PMA/ionomycin/Brefeldin A alone may not be sufficient for IL-5 production. Our testing method is as follows: Purified CD4+ T cells stimulated with immobilized anti-CD3 (10 μg/ml) + soluble anti-CD28 (2 μg/ml) + IL-2 (20 ng/ml) + IL-4 (50 ng/ml) for 2 days, wash and then culture in IL-2 + IL-4 for 3-4 days. You can find this information in our stimulation guide: https://www.biolegend.com/media_assets/support_protocol/BioLegend_StimulationGuide_101711.pdf

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