Suggested Staining Procedure for APC anti-Human GARP Antibody:
1. Harvest PBMC after stimulation (12 hours) with Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and Purified NA/LE
Mouse Anti-Human CD28 (Cat. No. 555725).
2. Wash the cells twice with stain buffer (eg. BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656).
3. Stain 1 X 10^6 cells either with the APC anti-Human GARP antibody (Cat. No. 562341) or with APC Mouse IgG2b, κ Isotype Control
(Cat. No. 555745) for 30 minutes on ice, protected from light.
4. Wash cells twice with stain buffer.
5. Stain for FoxP3. Refer to the Technical Data Sheet of BD Horizon™ V450 Mouse Anti-Human FoxP3, Cat. No. 560459, for detailed
FoxP3 staining protocol.
a. Add 2 ml of 1 X FoxP3 buffer A to the cell pellet.
b. Centrifuge and incubate in 0.5 ml of buffer C for 30 minutes.
c. Wash twice with stain buffer and stain with V450 Anti-Human FoxP3 antibody (Cat. No. 560459) for 30-45 minutes.
d. Wash twice with stain buffer and acquire on the Flow cyotmeter.
Note: BD Pharmingen™ Human FoxP3 Buffer Set (Cat. No. 560098) is a suggested companion product.