. 2021 Apr 5;2(2):100425. doi: 10.1016/j.xpro.2021.100425

Table1.

Preparation of the labeling mix

Reagent	Label	Dilution	Protocol
anti-CD31	BV421	1:100	Both
anti-CD45	BV421	1:100	Both
anti-TER119	BV421	1:200	Both
anti-O4	AF405	1:100	Only A^a
anti-O4	Biotinylated	1:100	Only B^a
Streptavidin	BV421	1:300	Only B^a
EGF	AF488	1:300	Both
anti-CD24	PerCP- Cy5.5	1:300	Only A^a
anti-CD24	BB700	1:300	Only B^a
anti-GLAST	PE	1:100	Both
anti-CD9	APC-Vio770	1:20	Both

In protocol B, antibodies to detect O4 and CD24 have been substituted because the signal from AF405 and PerCEP-Cy5.5 fluorochromes is greatly quenched as a result of fixation. Instead, the combination of biotinylated anti-O4 with BV421-Streptavidin and BB700-conjugated anti-CD24 provides a strong fixative-resistant signal using the same laser/filter cytometer configuration.