Product: Brilliant Violet 650™ anti-human IFN-γ

Catalog No.: 502538

Romy Hackbusch, Miss University Medical Center Hamburg-Eppendorf

Anti-human IFNg-Brilliant Violet 650™
Overall:	Product Quality:	Ease of Use:
You need a lot of antibody to detect IFNg.

Experimental Design

Application:	Flow Cytometry
Cells used:	Whole blood
Brief Protocol:	1. Stimulate whole blood with PMA/Ionomycin for 5 hours at 37°C, 5% CO2 2. Stain additional surface markers 3. Lyse erythrocytes 4. Wash two times with PBS, centrifuge at 400 x g for 5 minutes; discard supernatant 5. Permeabilisation with Saponin 6. staining of IFNg 7. Wash cells with PBS 8. Add 200-300µl FACS-buffer, FACS analysis The antibody was titrated and is used with 10µl per test
Results Summary:	Not the best antibody for staining IFNg.
Additional Notes:	Gating strategy: Lymphocytes > single cells > CD4+ cells > IFNg+

Data Image Click on the image for full size data.

Publication Status This data has not been published.

Technical Service Notes

Using nuclear factor permeabilization buffers for cytokine staining is generally not recommended as it can cause cytokines to leak from the cell. Even for a factor like IFN-g which can be in high abundance, a control sample (that did not permeabilize to the nucleus) is required in order to confirm no cytokine was lost.

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