The siPSCs were identified after typical colony formation. The differentiation ability of EB was measured on the 7th day in a 24-well cell culture plate. Cells were fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 for 10 min, and then blocked with 2% BSA for 30 min at room temperature. The primary antibodies against Sox2 (1：200, Abcam), Oct-3/4 (1：100, Santa Cruz), Nanog (1：100, Santa Cruz), Rex-1 (1：100, Santa Cruz), SSEA-1 (1：20, Developmental Studies Hybridoma Bank), E-cadherin (1：100, BD), Tra-1-60 (1：200, Millipore), Tra-1-81 (1：200, Millipore), smooth muscle actin (SMA, 1：200, Santa Cruz), β-tubulin (Tuj-1, 1：100, Santa Cruz), and human Sox17 (Sox17, 1：100, R&D) were added and incubated overnight at 4℃. Cy3-labeled anti-mouse IgG (1：500, Beyotime, China) and anti-mouse IgM (1：500, Bioss, China) secondary antibodies were added for 1 h at 37℃ for detection of target proteins. Nuclei were stained with DAPI (1：1,000, Sigma) for 5 min.
Rex 1
Manufactured by Santa Cruz Biotechnology
Rex-1 is a protein detection reagent used for western blot analysis. It is a rabbit polyclonal antibody that specifically binds to the Rex-1 protein, which is a transcription factor involved in the regulation of embryonic stem cell pluripotency.
Automatically generated - may contain errors
Lab products found in correlation
Tra 1 60, by Merck Group (2 mentions)
Tra 1 81, by Merck Group (2 mentions)
Nanog, by Santa Cruz Biotechnology (1 mentions)
Sox17, by R&D Systems (1 mentions)
Oct3 4, by Santa Cruz Biotechnology (1 mentions)
Ssea 1, by Developmental Studies Hybridoma Bank (1 mentions)
Tuj 1, by Santa Cruz Biotechnology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
E cadherin, by BD (1 mentions)
Lin28a, by Abcam (1 mentions)
Nanog, by Cell Signaling Technology (1 mentions)
Axio imager 2, by Zeiss (1 mentions)
2 protocols using rex 1
1
Immunofluorescence Characterization of siPSCs
2
Immunocytochemistry for Pluripotent Stem Cells
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In a 4-well plate, cover slips were coated with Matrigel for 30 min, 2 h or overnight; and then incubated with StemFlex medium for 30 min or 2 h to equilibrate the cover slips before cells were seeded on them. H1 and IMR90-1 cell lines and iPSCs from each selected clone that was chosen to be characterized for pluripotency were cultured onto cover slips. When colonies reached the appropriate size, which was usually about 4–5 days post passage, the immunocytochemistry procedures were performed as previously described42 (link) using appropriate primary (Oct4 (Rabbit, Stemgent), Sox2 (Rabbit, Stemgent), Nanog (Mouse, Cell Signaling), Klf4 (Rabbit, Abcam), Lin28A (Rabbit, Abcam), TRA-1-60 (Mouse, Millipore), TRA-1-81 (Mouse, Millipore) and Rex1 (Mouse, Santa Cruz) and secondary antibodies dilutions (Supplementary Information ). Cells were observed and photographed by a fluorescence microscope (Zeiss Axio Imager 2) using 40X objective.
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