Table 2.
Reagents details.
|
Antibodies used for immunocytochemistry/flow-cytometry |
||||
|---|---|---|---|---|
| Antibody | Dilution | Company Cat # | RRID | |
| Pluripotency Markers | Mouse anti-TRA1-60 | 1:200 | Cell Signaling Technology Cat#4746S | AB_2119059 |
| Rabbit anti-OCT4 | 1:100 | Thermo Fisher Scientific Cat#PA596860 | AB_2808662 | |
| Rabbit anti-SOX2 | 1:500 | Merck Millipore Cat#AB5603 | AB_2286686 | |
| Mouse anti-TRA1-81 | 1:200 | Cell Signaling Technology Cat#4745S | AB_2119060 | |
| Rabbit anti-NANOG | 1:500 | ThermoFisher Scientific Cat#PA1-097 | AB_2539867 | |
| Secondary antibodies |
AF555 Goat anti-Mouse, IgM | 1:500 | Thermo Fisher Scientific Cat#A21426 | AB_2535847 |
| AF488 Goat anti-Rabbit, IgG | 1:500 | Thermo Fisher scientific Cat#A11034 | AB_2576217 | |
|
Primers |
||||
| Target | Size of band | Forward/Reverse primer (5′-3′) | ||
| Pluripotency Markers (RT-qPCR) | DNMT3B | 55 bp | Hs00171876_m1 | |
| NANOG | 99 bp | Hs04260366_g1 | ||
| POU5F1 | 77 bp | Hs04260367_gH | ||
| SOX2 | 91 bp | Hs01053049_s1 | ||
| House-Keeping Genes (RT-qPCR) | GAPDH | 93 bp | Hs02758991_g1 | |
| ACTB | 63 bp | Hs01060665_g1 | ||
| Differentiation markers (RT-qPCR) | CXCR4 | 153 bp | Hs00607978_s1 | |
| FOXA2 | 66 bp | Hs00232764_m1 | ||
| SOX17 | 149 bp | Hs00751752_s1 | ||
| NKX2.5 | 64 bp | Hs00231763_m1 | ||
| αSMA (ACTA2) | 105 bp | Hs00426835_g1 | ||
| HAND1 | 54 bp | Hs00231848_m1 | ||
| HES5 | 62 bp | Hs01387463_g1 | ||
| MAP2 | 98 bp | Hs00258900_m1 | ||
| PAX6 | 76 bp | Hs00240871_m1 | ||
| Targeted mutation sequencing | BGN c.776G > T | 319 bp | GTTTTCCCAGTCACGACAAGGGTGATGCCAGAGTCC/ CAGGAAACAGCTATGACGACTGAGGGACTGCCCG | |
| Sendai virus Plasmids (PCR) | SeV | 181 bp | GGATCACTAGGTGATATCGAGC/ACCAGACAAGAGTTTAAGAGATATGTATC | |
| KOS | 528 bp | ATGCACCGCTACGACGTGAGCGC/ ACCTTGACAATCCTGATGTGGyc | ||
| Klf4 | 410 bp | TTCCTGCATGCCAGAGGAGCCC/AATGTATCGAAGGTGCTCAA | ||
| c-Myc | 532 bp | TAACTGACTAGCAGGCTTGTCG/ TCCACATACAGTCCTGGATGATGATG | ||