Table 2.
Reagents details.
| Antibodies used for immunocytochemistry/flow-citometry | |||
|---|---|---|---|
| Antibody | Dilution | Company Cat # and RRID | |
| Pluripotency Markers (Immunocytochemistry) | Rabbit anti-OCT4 | 1:100 | Thermo Fisher Scientific Cat# A24867, RRID:AB_2650999 |
| Pluripotency Markers (Immunocytochemistry) | Mouse anti-SSEA4 (IgG3) | 1:100 | Thermo Fisher Scientific Cat# A24866, RRID:AB_2651001 |
| Pluripotency Markers (Immunocytochemistry) | Rat anti-SOX2 | 1:100 | Thermo Fisher Scientific Cat# A24759, RRID:AB_2651000 |
| Pluripotency Markers (Immunocytochemistry) | Mouse anti-TRA-1-60 (IgM) | 1:100 | Thermo Fisher Scientific Cat# A24868, RRID:AB_2651002 |
| Secondary antibodies (Immunocytochemistry) | Alexa Fluor® 555 Donkey Anti-Rabbit | 1:250 | Thermo Fisher Scientific Cat# A24869, RRID:AB_2651006 |
| Secondary antibodies (Immunocytochemistry) | Alexa Fluor® 488 Goat Anti-Mouse IgG3 | 1:250 | Thermo Fisher Scientific Cat# A24877, RRID:AB_2651008 |
| Secondary antibodies (Immunocytochemistry) | Alexa Fluor® 488 Donkey Anti-Rat | 1:250 | Thermo Fisher Scientific Cat# A24876, RRID:AB_2651007 |
| Secondary antibodies (Immunocytochemistry) | Alexa Fluor® 555 Goat Anti-Mouse IgM | 1:250 | Thermo Fisher Scientific Cat# A24871, RRID:AB_2651009 |
| Pluripotency Markers (Flow cytometry) | PE-SOX2 Mouse IgG2A | 1:20 | R&D System Cat# IC2018P, RRID:AB_357273 |
| Pluripotency Markers (Flow cytometry) | CFS-SSEA-4 Mouse IgG3 | 1:20 | R&D System Cat# FAB1435F, RRID:AB_952015 |
| Pluripotency Markers (Flow cytometry) | PE Isotype control- mouse IgG2A | 1:20 | R&D System Cat# IC003P, RRID:AB_357245 |
| Pluripotency Markers (Flow cytometry) | CFS Isotype control- Mouse IgG3 | 1:20 | R&D System Cat# IC007F, RRID:AB_952037 |
| Cardiomyocyte markers (Immunocytochemistry) |
Mouse anti-Sarcomeric actinin | 1:250 | Sigma Aldrich Cat# A7732, RRID:AB_2221571 |
| Cardiomyocyte markers (Immunocytochemistry) |
Rabbit anti-Troponin I (H-170) | 1:500 | Santa Cruz Biotechnology Cat# sc-15,368, RRID:AB_793465 |
| Secondary antibodies (Immunocytochemistry) |
Alexa Fluor® 488 Goat Anti-Mouse IgG | 1:1000 | Thermo Fisher Scientific Cat# A11029, RRID:AB_2534088 |
| Secondary antibodies (Immunocytochemistry) |
Alexa Fluor® 555 Goat Anti-Rabbit IgG | 1:1000 | Thermo Fisher Scientific Cat# A21429, RRID:AB_2535850 |
| Primers | ||
|---|---|---|
| Target | Forward/Reverse primer (5′-3′) | |
| Pluripotency Markers (qRT-PCR) | SOX2 | GGGAAATGGGAGGGGTGCAAAAGAGG/TTGCGTGAGTGTGGATGGGATTGGTG |
| Pluripotency Markers (qRT-PCR) | OCT4 | GACAGGGGGAGGGGAGGAGCTAGG/CTTCCCTCCAACCAGTTGCCCCAAAC |
| Pluripotency Markers (qRT-PCR) | NANOG | TGCAAGAACTCTCCAACATCCT/ATTGCTATTCTTCGGCCAGTT |
| Three germ layer markers (endoderm) (qRT-PCR) | SOX7 | TGAACGCCTTCATGGTTTG/AGCGCCTTCCACGACTTT |
| Three germ layer markers (endoderm) (qRT-PCR) | AFP | GTGCCAAGCTCAGGGTGTAG/CAGCCTCAAGTTGTTCCTCTG |
| Three germ layer markers (mesoderm) (qRT-PCR) | CD31 | ATGCCGTGGAAAGCAGATAC/CTGTTCTTCTCGGAACATGGA |
| Three germ layer markers (mesoderm) (qRT-PCR) | ACTA2 | GTGATCACCATCGGAAATGAA/TCATGATGCTGTTGTAGGTGGT |
| Three germ layer markers (mesoderm) (qRT-PCR) | SCL | CCAACAATCGAGTGAAGAGGA/CCGGCTGTTGGTGAAGATAC |
| Three germ layer markers (mesoderm) (qRT-PCR) | CDH5 | GAGCATCCAGGCAGTGGTAG/CAGGAAGATGAGCAGGGTGA |
| Three germ layer markers (ectoderm) (qRT-PCR) | KRT14 | CACCTCTCCTCCTCCCAGTT/ATGACCTTGGTGCGGATTT |
| Three germ layer markers (neuro-ectoderm) (qRT-PCR) | NCAM1 | CAGATGGGAGAGGATGGAAA/CAGACGGGAGCCTGATCTCT |
| Three germ layer markers (neuro-ectoderm) (qRT-PCR) | GABRR2 | CTGTGCCTGCCAGAGTTTCA/ACGGCCTTGACGTAGGAGA |
| House-Keeping Gene (qRT-PCR) | GAPDH | CCACCCATGGCAAATTCC/TCGCTCCTGGAAGATGGTG |
| Episomal plasmid Gene (episome silencing) | EBNA-1 | ATCAGGGCCAAGACATAGAGATG/GCCAATGCAACTTGGACGTT |
| House-Keeping Gene (episome silencing) | FBXO15 | GCCAGGAGGTCTTCGCTGTA/AATGCACGGCTAGGGTCAAA |
| Mutation analysis (PCR) | PKP2 | GAACACCCACAGGCCGC/TTTCTTGGGCTGGGTAGTAGAAA |
| Mutation analysis (sequencing) | PKP2 | GCCTCACTCATTCTCCCTGATCTCAG/TAGGCTTTGGCAGTCCGGCTGTTG |