Table 2.
Reagents details.
| Antibodies Used for Immunocytochemistry | |||
|---|---|---|---|
| Antibody | Dilution | Company Cat # and RRID | |
| Pluripotency Markers | Rabbit IgG anti-OCT4 | 1:200 | Abcam, Cambridge, UK, Cat # ab18976, RRID:AB_444714 |
| Mouse IgG3 anti-SSEA4 | 1:200 | Abcam, Cambridge, UK, Cat # ab16287, RRID:AB_778073 | |
| Mouse IgM anti-TRA-1-60 | 1:200 | Abcam, Cambridge, UK, Cat # ab16288, RRID:AB_778563 | |
| Rabbit IgG anti-SOX2 | 1:500 | Cell Signaling, Danvers, MA, USA, Cat # 3579, RRID:AB_2195767 | |
| Differentiation Markers | Mouse IgG2a anti-αSMA | 1:100 | Dako, Glostrup, Denmark, Cat # M0851, RRID:AB_2223500 |
| Rabbit IgG anti-NKX2.5 (H-114) |
1:100 | Santa Cruz Biotechnology, Dallas, TX, USA, Cat # sc-14033, RRID:AB_650281 | |
| Mouse IgG2a anti-AFP | 1:250 | Sigma-Aldrich, Darmstadt, Germany, Cat # A8452, RRID:AB_258392 | |
| Mouse IgG2a anti-Tubulin β 3 (TUBB3)/Clone: TUJ1 | 1:1000 | BioLegend, San Diego, CA, USA, Cat # 801,201, RRID:AB_2313773 | |
| Chicken IgG anti MAP2 | 1:1000 | Abcam, Cambridge, UK, Cat # ab5392, RRID:AB_2138153 | |
| Mouse IgG1 anti-HNF3b (FOXA2) | 1:50 | Santa Cruz Biotechnology, Dallas, TX, USA, Cat # sc-374,376, RRID:AB_10989742 | |
| Mouse IgG1 anti-S100β | 1:400 | Sigma-Aldrich, Darmstadt, Germany, Cat # S2532, RRID:AB_477499 | |
| Rabbit IgG anti-GFAP | 1:200 | Dako, Glostrup, Denmark, Cat # Z0334 | |
| Secondary antibodies | Goat anti-Mouse IgG (H + L) Secondary Antibody, Alexa Fluor 488 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat # A11029, RRID:AB_2534088 |
| Goat anti-Mouse IgG (H + L) Secondary Antibody, Alexa Fluor 568 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat # A11031, RRID:AB_144696 | |
| Goat anti-Rabbit IgG (H + L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat # A11008, RRID:AB_143165 | |
| Goat anti-Rabbit IgG (H + L) Alexa Fluor 568 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat # A11011, RRID:AB_143157 | |
| Goat anti-Mouse IgG1 Alexa Fluor 568 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat # A21124, RRID:AB_2535766 | |
| Goat anti-Mouse IgG2a Alexa Fluor 488 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat # A21131, RRID:AB_2535771 | |
| Goat anti-Chicken IgY (H + L) Alexa Fluor 488 | 1:400 | Abcam, Cambridge, UK, Cat # ab150173, RRID:AB_2827653 | |
| Primers | |||
| Target | Size of band | Forward/Reverse primer (5′-3′) | |
| Episomal plasmid vector detection | EBNA-1 | 61 bp | TTCCACGAGGGTAGTGAACC/ TCGGGGGTGTTAGAGACAAC |
| Mycoplasma detection | 16S ribosomal RNA gene | 280 bp | GGGAGCAAACAGGATTAGATACCCT/ TGCACCATCTGTCACTCTGTTAACCTC |
| House-keeping gene (RT-qPCR) | beta-2-microglobulin | 280 bp | TAGCTGTGCTCGCGCTACT/ TCTCTGCTGGATGACGTGAG |
| Pluripotency marker (RT-qPCR) | NANOG | 116 bp | TTTGTGGGCCTGAAGAAAACT/ AGGGCTGTCCTGAATAAGCAG |
| OCT4 | 94 bp | CTTCTGCTTCAGGAGCTTGG/ GAAGGAGAAGCTGGAGCAAA |
|
| SOX2 | 100 bp | GCTTAGCCTCGTCGATGAAC/ AACCCCAAGATGCACAACTC |
|
| Targeted mutation analysis | GBA1 | 600 bp | CTGTTGCTACCTAGTCACTTCC/ CCCTATCTTCCCTTTCCTTCAC |