Table 2.
Reagents details.
| Antibodies Used for Immunocytochemistry | |||
|---|---|---|---|
| Antibody | Dilution | Company Cat # and RRID | |
| Pluripotency Markers | Mouse IgG2b anti-OCT3/4 (C-10) | 1:200 | Santa Cruz Biotechnology, Dallas, TX, USA, Cat# sc-5279, RRID:AB_628051 |
| Mouse IgG3 anti-SSEA-4 | 1:200 | Abcam, Cambridge, UK, Cat# ab16287, RRID:AB_778073 | |
| Mouse IgG1 anti-NANOG | 1:200 | Santa Cruz Biotechnology, Dallas, TX, USA, Cat# sc-293121, RRID:AB_2665475 | |
| 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 |
| Mouse IgG1 anti-CD29 (Integrin beta 1) (TS2/16) | 1:100 | Thermo Fisher Scientific, Waltham, MA, USA, Cat # 14-0299-82, RRID:AB_1210468 | |
| 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# 801201, RRID:AB_2313773 | |
| Chicken IgG anti MAP2 | 1:1000 | Abcam, Cambridge, UK, Cat# ab5392, RRID:AB_2138153 | |
| Mouse IgG1 anti-CK18 | 1:200 | Millipore, Burlington, VT, USA Cat# MAB3234, RRID:AB_94763 | |
| Macrophage-specific Markers | Mouse IgG2b, κ anti-CD14 APC (Clone MφP9) | 1:30 | BD Biosciences, Franklin Lakes, NJ, USA, Cat# 345787, RRID:AB_2868813 |
| Mouse IgG1, κ anti-CD45 PerCP-Cy5.5 CE | 1:20 | BD Biosciences, Franklin Lakes, NJ, USA, Cat# 332784, RRID:AB_2868632 | |
| Secondary antibodies | Goat anti-mouse IgG3 cross-adsorbed secondary antibody, Alexa Fluor 488 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat# A-21151, RRID:AB_2535784 |
| Goat anti-mouse IgG2b cross-adsorbed secondary Antibody, Alexa Fluor 568 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat# A-21144, RRID:AB_2535780 | |
| Goat anti-rabbit IgG (H + L) Alexa Fluor 568 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat# A-11011, RRID:AB_143157 | |
| Goat anti-mouse IgG1 Alexa Fluor 488 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat# A-21121, RRID:AB_2535764 | |
| Goat anti-mouse IgG1 Alexa Fluor 568 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat# A21124, RRID:AB_2535766 | |
| Goat anti-mouse IgG2a cross-adsorbed secondary antibody, Alexa Fluor 568 | 1:400 | Thermo Fisher Scientific, Waltham, MA, USA, Cat # A-21134, RRID:AB_2535773 | |
| 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 vectors detection | EBNA-1 | 61 bp | TTCCACGAGGGTAGTGAACC/ TCGGGGGTGTTAGAGACAAC |
| Mycoplasma detection | 16S ribosomal RNA gene | 280 bp | GGGAGCAAACAGGATTAGATACCCT/ TGCACCATCTGTCACTCTGTTAACCTC |
| House-keeping gene (RT-qPCR) | ACTB | 93 bp | GCACAGAGCCTCGCCTT/ GTTGTCGACGACGAGCG |
| Pluripotency marker (RT-qPCR) | NANOG | 116 bp | TTTGTGGGCCTGAAGAAAACT/ AGGGCTGTCCTGAATAAGCAG |
| OCT4 | 94 bp | CTTCTGCTTCAGGAGCTTGG/ GAAGGAGAAGCTGGAGCAAA |
|
| SOX2 | 100 bp | GCTTAGCCTCGTCGATGAAC/ AACCCCAAGATGCACAACTC |
|
| Targeted mutation analysis | MEFV | 297 bp | TGGGATCTGGCTGTCACATTG/ CATTGTTCTGGGCTCTCCGAG |