Product: Alexa Fluor® 647 anti-human FOXP3

Catalog No.: 320214

Romy Hackbusch, Miss University Medical Center Hamburg-Eppendorf

Anti-human FOXP3-Alexa Fluor® 647
Overall:	Product Quality:	Ease of Use:
Staining of FOXP3 works well but a lot of antibody is used.

Experimental Design

Application:	Flow Cytometry
Cells used:	Whole blood
Brief Protocol:	1. Stimulate whole blood with PMA/Ionomycin for 5 hours, 37°C, 5% CO2 2. Stain additional surface antibodies 3. Lysis of erythrocytes by adding 1ml of Lysis buffer, mix well, incubate for 10 minutes at RT, dark. 4. Wash two times with PBS; centrifuge at 400 x g for 5 minutes; trash supernatant 5. Fixation/Permeabilisation with FoxP3 Buffer Set, incubate 45 minutes, RT 6. Wash with Permeabilisation-buffer; centrifuge at 400 x g for 5 minutes; trash supernatant 7. Stain FoxP3 ; incubate 30 minutes, RT, dark 8. Wash with Permeabilisation-buffer; centrifuge at 400 x g for 5 minutes; trash supernatant 9. Add 200-300µl FACS-Buffer, FACS analysis 10. This antibody was titrated and is use with a amount of 10µl/test
Results Summary:	High amount of antibody is needed but staining works well.
Additional Notes:	Gating strategy: Lymphocytes > single cells > CD4+ cells > FOXP3+ cells

Data Image Click on the image for full size data.

Publication Status This data has not been published.

Technical Service Notes

True-Nuclear Transcription Factor Buffer is recommended for all FOXP3 antibodies to improve staining: http://www.biolegend.com/true-nuclear-transcription-factor-buffer-set-10859.html

See all Reviews