Table 3:
Reagents details
| Antibodies used for immunocytochemistry/flow-cytometry | |||
|---|---|---|---|
| Antibody | Dilution | Company Cat # and RRID | |
| Self-renewal Markers | Rabbit anti-NANOG | 1:200 | Stemgent Cat# 09–0020, RRID:AB_2298294 |
| Self-renewal Markers | Mouse anti-human OCT-3/4 | 1:1000 | Santa Cruz Biotechnology Cat# sc-5279, RRID:AB_628051 |
| Self-renewal Markers | Goat anti-human SOX2 | 1:1000 | R&D Systems Cat# AF2018, RRID:AB_355110 |
| Secondary antibodies | cy5 Donkey anti-rabbit | 1:500 | Jackson ImmunoResearch Labs Cat# 711–175-152, RRID:AB_2340607 |
| Secondary antibodies | 488 donkey anti-mouse | 1:500 | Molecular Probes Cat# A-21202, RRID:AB_141607 |
| Secondary antibodies | Cy3 donkey anti-goat | 1:500 | Jackson ImmunoResearch Labs Cat# 705–165-147, RRID:AB_2307351 |
| Primers | |||
| Target | Forward/Reverse primer (5′−3′) | ||
| Episomal plasmids | All episomal plasmids pCXLE-hSK 2.8 kB pCXLE-hUL 2.1 kB pCXLE-hOCT3/4-shp53 1.4 kB |
GTGACCGGCGGCTCTAGAGC/AAGCCATACGGGAAGCAATAGC | |
| gDNA control | Positive DNA control for episomal PCR (1.3 kB) | CATGCAGTCCTCCTTACCATC TCCTCTCTGGCTCCATCGTA | |
| Trilineage potential - experiment controls |
ACTIN B Loading Control GAPDH Loading Control |
Qiagen primer set number: PPH00073G PPH00150F |
|
| Trilineage potential – Ectodermal genes |
PAX6 NR2F2 |
PPH02598F PPH05883B |
|
| Trilineage potential – Mesodermal genes |
HAND1 MIXL1 |
PPH06879B PPH15840A |
|
| Trilineage potential – Endodermal genes |
GATA6 SOX17 |
PPH06943F PPH02451A |
|