Materials
Name | Company | Catalog Number | Comments |
pCX-OKS-2A pDNA | Addgene | 19771 | Obtained from Addgene as bacterial stab, plasmid production performed in Plasmid Factory (Germany) |
pCX-cMyc | Addgene | 19772 | Obtained from Addgene as bacterial stab, plasmid production performed in Plasmid Factory (Germany) |
0.22 μm filter | Milliopore | SLGP033RS | |
Isoflurane | Abbott | B505 | |
HBSS buffer | Sigma-Aldrich | H6648 | Ca2+ and Mg2+ free, with bicarbonate |
Liver Digest Medium | Gibco | 17703-034 | |
Hepatocyte Wash Medium | Gibco | 17704-024 | |
100 μm cell strainer | BD Biosciences | 352360 | |
Nucleospin RNA II kit | Macherey-Nagel | 740955.25 | Kit for RNA isolation from cells and tissues |
iScript cDNA synthesis kit | Bio-Rad | 170-8890 | |
iO SYBR Green Supermix | Bio-Rad | 170-8880 | |
Oct3/4 forward primer | Sigma | sequence given in comments | TGAGAACCTTCAGGAGATATGCAA |
Oct3/4 reverse primer | Sigma | sequence given in comments | CTCAATGCTAGTTCGCTTTCTCTTC |
Sox2 forward primer | Sigma | sequence given in comments | GGTTACCTCTTCCTCCCACTCCAG |
Sox2 reverse primer | Sigma | sequence given in comments | TCACATGTGCGACAGGGGCAG |
C-myc forward primer | Sigma | sequence given in comments | CAGAGGAGGAACGAGCTGAAGCGC |
C-myc reverse primer | Sigma | sequence given in comments | TTATGCACCAGAGTTTCGAAGCTGTTCG |
Nanog forward primer | Sigma | sequence given in comments | CAGAAAAACCAGTGGTTGAAGACTAG |
Nanog reverse primer | Sigma | sequence given in comments | GCAATGGATGCTGGGATACTC |
Ecat1 forward primer | Sigma | sequence given in comments | TGTGGGGCCCTGAAAGGCGAGCTGAGAT |
Ecat1 reverse primer | Sigma | sequence given in comments | ATGGGCCGCCATACGACGACGCTCAACT |
Rex1 forward primer | Sigma | sequence given in comments | ACGAGTGGCAGTTTCTTCTTGGGA |
Rex1 reverse primer | Sigma | sequence given in comments | TATGACTCACTTCCAGGGGGCACT |
Alb forward primer | Sigma | sequence given in comments | GTTCGCTACACCCAGAAAGC |
Alb reverse primer | Sigma | sequence given in comments | CCACACAAGGCAGTCTCTGA |
Aat forward primer | Sigma | sequence given in comments | CAGAGGAGGCCAAGAAAGTG |
Aat reverse primer | Sigma | sequence given in comments | ATGGACAGTCTGGGGAAGTG |
Trf forward primer | Sigma | sequence given in comments | ACCATGTTGTGGTCTCACGA |
Trf reverse primer | Sigma | sequence given in comments | ACAGAAGGTCCTTGGTGGTG |
B actin forward primer | Sigma | sequence given in comments | GACCTCTATGCCAACACAGT |
B actin reverse primer | Sigma | sequence given in comments | AGTACTTGCGCTCAGGAGGA |
BD Cytofix fixation buffer | BD Biosciences | 560585 | |
1x BD Perm/Wash buffer | BD Biosciences | 560585 | |
anti-mouse OCT4-PerCP-Cy5.5 | BD Biosciences | 560585 | |
anti-mouse Nanog-PE | BD Biosciences | 560585 | |
2-Methylbutane | Sigma-Aldrich | M32631 | |
X100-Triton | Sigma-Aldrich | X100-500ML | |
Goat serum | Sigma-Aldrich | G9023 | |
BSA | Sigma | A2153 | |
rabbit pAb anti-OCT4 | Abcam | ab19857 | Use at a concentration of 3 μg/ml |
rabbit pAb anti-SOX2 | Abcam | ab97959 | Use at a concentration of 1 μg/ml |
rabbit pAb anti-Nanog | Abcam | ab80892 | Use at a concentration of 1 μg/ml |
mouse mAb anti-SSEA1 | Abcam | ab16285 | Use at a concentration of 20 μg/ml |
goat pAb anti-rabbit IgG labeled with Cy3 | Jackson ImmunoResearch Laboratories Inc. | 111-165-003-JIR | Use at a concentration of 1/250 |
goat pAb anti-mouse IgG labeled with Cy3 | Jackson ImmunoResearch Laboratories Inc. | 115-165-003-JIR | Use at a concentration of 1/250 |
VECTASHIELD mounting medium with DAPI | Vector Laboratories | H-1200 | |
BCIP/NBT liquid substrate system | Sigma | B1911 | |
Aqueous mounting medium | Sigma | ||
16% Paraformaldehyde | Fisher | AA433689M | Stock solution is 16%, use it at 4% |
Hematoxylin and eosin | Sigma | GH53/HT110216 | |
PAS staining | Sigma | 395B |
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