Table 3.
Reagents details.
| Antibodies used for immunocytochemistry/flow-citometry | |||
|---|---|---|---|
| Antibody | Dilution | Company Cat # and RRID | |
| Pluripotency Markers | Rabbit anti-NANOG | 1:200 | Cell Signaling Technology Cat# 4903, RRID:AB_10559205 |
| Rabbit anti-Sox2 | 1:100 | Abcam Cat# ab15830, RRID:AB_443255 | |
| Rabbit anti-Pou5f1 | 1:100 | Santa Cruz Biotechnology Cat# sc-9081, RRID:AB_2167703 | |
| Differentiation Markers | Mouse Anti-AFP | 1:100 | Sigma-Aldrich Cat# A8452, RRID:AB_258392 |
| Rabbit Anti- αSMA | 1:100 | Abcam Cat# ab124964, RRID:AB_11129103 | |
| Mouse Anti- TUBB3 | 1:100 | BioLegend Cat# 801202, RRID:AB_10063408 | |
| Secondary antibodies | Alexa flour 594 donkey anti rabbit | 1:1000 | Thermo Fisher Scientific Cat# A-21207, RRID:AB_141637 |
| Alexa flour 488 donkey anti rabbit | 1:1000 | Thermo Fisher Scientific Cat# A-21206, RRID:AB_2535792 | |
| Alexa flour 594 goat anti mouse | 1:1000 | Thermo Fisher Scientific Cat# A-21203, RRID:AB_2535789 | |
| Thermo Fisher Scientific Cat# A-21203, RRID:AB_2535789 | |||
| Primers | |||
| Target | Forward/Reverse primer (5′-3′) | ||
| Genotyping | PCCB | Fwd-120:GGAAAGCTAACAGCCAG/Rev120:CTTATTCTTATCAGCAGC | |
| Fwd131:TTCCATTGTAGGGAAAGC/Rev131:CTTATTCTTATCAGCAGC | |||
| Targeted mutation analysis/sequencing | PCCB | Fwd-XbaI:TTTTCTAGAGGAAAGCTAACAGCCAG/Rev-EcoRI:AACGAATTCCTTATTCTTATCAGCAGC | |
| Fwd-XbaI:TTTTCTAGATTCCATTGTAGGGAAAGA//Rev-EcoRI:AACGAATTCCTTATTCTTATCAGCAGC | |||
| Fwd615: CTTCCACAGTTGGGTGGCT /Rev615: AAGGCAGGGTACGGTCGCT | |||
| Fwd1230: CACTGTCAGGTGTGGGGTCCCT/Rev1230: ATGAGTTTGGCCACTATGCCTCA | |||