Table 2.
Antibodies used for immunocytochemistry/flow-cytometry | |||
---|---|---|---|
Antibody | Dilution | Company Cat # and RRID | |
Pluripotency Marker | Rabbit mAb anti-OCT4A | 1:400 | StemLight kit, Cell Signaling Technology, Cat# 9092 |
Pluripotency Marker | Rabbit mAb anti-SOX2 | 1:400 | StemLight kit, Cell Signaling Technology, Cat# 9092 |
Pluripotency Marker | Rabbit mAb anti-NANOG | 1:400 | StemLight kit, Cell Signaling Technology, Cat# 9092 |
Pluripotency Marker | Rabbit mAb anti-LIN28A | 1:400 | StemLight kit, Cell Signaling Technology, Cat# 9092 |
Pluripotency Marker | PE-conjugated mouse mAb anti-SSEA4 | 1:20 | ThermoFisher, Cat# 12-8843-41 |
Secondary antibody | Goat anti-Rabbit IgG Alexa Fluor Plus 488 | 1:500 | ThermoFisher, Cat# A32731 |
Isotype control antibody | Mouse IgG3 Isotype Control, PE | 1:20 | ThermoFisher, Cat# 12-4742-42 |
Primers | |||
Target | Forward/Reverse primer (5′-3′) | ||
Mycoplasma detection | Mycoplasma 16S rRNA |
GGCGAATGGGTGAGTAACACG CGGATAACGCTTGCGACCTATG |
|
POGLUT1 mutation fragment amplification | A 506-bp amplicon containing mutation region |
TGACCTGAACAACATACCCTTCA GCTAATGCTGGTTCATGGAACTT |
|
POGLUT1 amplicon sequencing | A 20-bp region located 125-bp upstream of the mutation within the amplicon | GTCCTAGTCCTGCTCACCTT |