Table 2:
Reagents details
| Antibodies used for immunocytochemistry/flow cytometry | ||||
|---|---|---|---|---|
| Antibody | Dilution | Company Cat # | RRID | |
| Pluripotency Markers | Mouse anti-Oct3/4 (C-10) | 1:200 | Santa Cruz #sc-5297 | RRID:AB_628051 |
| Rabbit anti-Nanog (D73G4) | 1:400 | Cell Signaling #4903S | RRID:AB_10559205 | |
| Rabbit anti-Sox2 (D6D9) | 1:300 | Cell Signaling #3579S | RRID:AB_2195767 | |
| AF488 anti-human SSEA-3 | 1:50 | BioLegend #330306 | RRID:AB_1279440 | |
| AF647 anti-human SSEA-4 | 1:400 | BioLegend #330408 | RRID:AB_1089200 | |
| AF488 anti-human Tra-1-60 | 1:100 | BioLegend #330614 | RRID:AB_2119064 | |
| AF647 anti-human Tra-1-81 | 1:50 | BioLegend #330706 | RRID:AB_1089242 | |
| Differentiation Markers | PE mouse anti-human Sox17 | 1:25 | BD #561591 | RRID:AB_10717121 |
| Mouse anti-human FoxA2 | 1:100 | Santa Cruz #sc-101060 | RRID:AB_1124660 | |
| Rabbit anti-FOXG1 | 1:300 | Abcam #196868 | RRID:AB_2892604 | |
| AF647 anti-human PAX6 | 1:20 | BD #562249 | RRID:AB_2644844 | |
| PE/Cyanine7 anti-human CD41 | 1:400 | BioLegend #303718 | RRID:AB_10899413 | |
| APC Mouse anti-human CD235 | 1:5000 | BD #551336 | RRID:AB_398499 | |
| Secondary Antibodies | Goat anti-mouse IgG2a-AF647 | 1:400 | Jackson Immunoresearch #115-605-206 | RRID:AB_2338917 |
| Goat anti-rabbit IgG-AF488 | 1:400 | Jackson Immunoresearch #111-545-144 | RRID: AB_2338052 | |
| Goat anti-mouse IgG2b-AF488 (flow cytometry) | 1:400 | Jackson Immunoresearch #115-545-207 | RRID:AB_2338856 | |
| Goat anti-Mouse IgG (H+L)-AF488 (immunohistochemistry) | 1:400 | ThermoFisher #A-11029 | RRID:AB_2534088 | |
| Primers | ||||
| Target | Size of band |
Forward/Reverse primer (5′-3′) | ||
| Sendai Screening (Taqman qRT-PCR) | SEV | 59 | Mr04269880_mr | |
| SEV-KLF4 | 67 | Mr04421256_mr | ||
| SEV-KOS | 80 | Mr04421257_mr | ||
| SEV-cMYC | 89 | Mr04269876_mr | ||
| GAPDH | 157 | Hs02786624_g1 | ||
| Targeted mutation analysis/sequencing | GATA1 Exon 2 screen | 387 bp | AGATGCAGGAGGGAAAAGAG / CGGCACATCCATTTGAGAAG | |
| GATA1 sequencing | AAGAGGAGCAGGTGAA | |||
| Mycoplasma Detection | 16S Ribosomal RNA | 518 bp | CGCCTGAGTAGTACGTTCGC / GCGGTGTGTACAAGACCCGA | |
| GAPDH (internal control) | 150 bp | GTGGACCTGACCTGCCGTCT / GGAGGAGTGGGTGTCGCTGT | ||