Table 3.
Reagents details. RRID Requirement for antibodies: use http://antibodyregistry.org/ to retrieve RRID for antibodies and include ID in table as shown in examples.
| Antibodies used for immunocytochemistry/flow-cytometry | |||
|---|---|---|---|
| Antibody | Dilution | Company Cat # and RRID | |
| Pluripotency Markers | Mouse anti-SOX2 | 1:250 | BioLegend, Cat# 656102, RRID: AB_2,562246 |
| Pluripotency Markers | Rabbit anti-NANOG | 1:400 | Cell Signaling Technology, Cat# 4903, RRID: AB_10559205 |
| Pluripotency Markers | Rabbit anti-OCT4 | 1:400 | Thermo Fisher, Cat# 701756, RRID: AB_2633031 |
| Pluripotency Markers | Mouse anti-SSEA4 | 1:1000 | Cell Signaling Technology, Cat# 4755, RRID: AB_1264259 |
| Secondary Antibodies | Donkey anti-Mouse IgG (Alexa Fluor 488) | 1:400 | Thermo Fischer, Cat# A21202, RRID: AB_141607 |
| Secondary Antibodies | Donkey anti-Rabbit IgG (Alexa Fluor 594) | 1:400 | Thermo Fischer, Cat# A21207, RRID: AB_141637 |
| Flow Cytometry Antibodies | Anti-Tra-1-60-DyLight 488 | 1:50 | Thermo Fischer, Cat# MA1–023-D488X, RRID: AB_2536700 |
| Flow Cytometry Antibodies | Anti-Nanog-Alexa Fluor 488 | 1:50 | Millipore, Cat# FCABS352A4, RRID: AB_10807973 |
| Flow Cytometry Antibodies | Anti-SSEA-4-Alexa Fluor 488 | 1:50 | Thermo Fischer, Cat# 53-8843-41, RRID: AB_10597752 |
| Flow Cytometry Antibodies | Mouse-IgM-DyLight 488 | 1:50 | Thermo Fischer, Cat# MA1-194-D488, RRID: AB_2536969 |
| Flow Cytometry Antibodies | Rabbit IgG-Alexa Fluor 488 | 1:50 | Cell Signaling Technology, Cat# 4340S, RRID: AB_10694568 |
| Flow Cytometry Antibodies | Mouse IgG3-FITC | 1:50 | Thermo Fischer, Cat# 11-4742-42, RRID: AB_2043894 |
| Primers | |||
| Target | Forward/Reverse primer (5′ – 3′) | ||
| Mycoplasma detection primers (qPCR) | GPO-1_MGSO/724bp | 5′-ACGGCCCAGACTCCTACGGGAGGCAGCAGTA | |
| 5′-CCATGCACCATCTGTCACTCTGTTAACCTC | |||
| House-keeping gene primers (qPCR) | GAPDH-3/488 bp | 5′-GGGAGCCAAAAGGGTCATCA | |
| 5′-TGATGGCATGGACTGTGGTC | |||