IPSC lines were rinsed with 1× PBS, fixed with 4% paraformaldehyde, and stored in PBS for staining. Cells were permeabilized with PBST (PBS supplemented with 0.1% Triton X-100) for 10 min at room temperature. Non-specific antigens were then blocked by incubating the cells with PBST+BSA at room temperature for 1 hr. Cells were incubated overnight at 4°C in PBS-BSA containing 10% donkey serum and the following specific primary antibodies: 1:500 anti-human OCT4 (Cell Signaling Technologies), 1:500 anti-human NANOG (Cell Signaling Technologies), 1:500 anti-human SSEA4 (Cell Signaling Technologies),), 1:500 anti-TRA-1–60 (Cell Signaling Technologies), 1:500 anti-SOX2 (Cell Signaling Technologies), and 1:200 anti-Lin28 (R&D Systems). Following 3 washes with PBS, cells were incubated with appropriate Alexa Fluor conjugated secondary antibodies at 1:200 dilution. After 3 washes with PBS, the samples were incubated for 10 min with DAPI (1 µg/mL) in PBS, followed by a final wash in PBS. Fluorescence images were captured at 10× or 20× on a Zeiss LSM 710/780 confocal microscope and processed using ImageJ.
Anti human oct4
Manufactured by Cell Signaling Technology
Anti-human OCT4 is a primary antibody that specifically recognizes the OCT4 protein, a transcription factor essential for the maintenance of embryonic stem cell pluripotency.
Automatically generated - may contain errors
Lab products found in correlation
Anti human ssea4, by Cell Signaling Technology (2 mentions)
Anti lin28, by R&D Systems (2 mentions)
Lsm 710 780 confocal microscope, by Zeiss (2 mentions)
Anti human nanog, by Cell Signaling Technology (2 mentions)
Anti tra 1 60, by Cell Signaling Technology (2 mentions)
Anti sox2, by Cell Signaling Technology (2 mentions)
2 protocols using anti human oct4
1
Characterization of Induced Pluripotent Stem Cells
2
Characterization of Induced Pluripotent Stem Cells
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IPSC lines were rinsed with 1× PBS, fixed with 4% paraformaldehyde, and stored in PBS for staining. Cells were permeabilized with PBST (PBS supplemented with 0.1% Triton X-100) for 10 min at room temperature. Non-specific antigens were then blocked by incubating the cells with PBST+BSA at room temperature for 1 hr. Cells were incubated overnight at 4°C in PBS-BSA containing 10% donkey serum and the following specific primary antibodies: 1:500 anti-human OCT4 (Cell Signaling Technologies), 1:500 anti-human NANOG (Cell Signaling Technologies), 1:500 anti-human SSEA4 (Cell Signaling Technologies),), 1:500 anti-TRA-1–60 (Cell Signaling Technologies), 1:500 anti-SOX2 (Cell Signaling Technologies), and 1:200 anti-Lin28 (R&D Systems). Following 3 washes with PBS, cells were incubated with appropriate Alexa Fluor conjugated secondary antibodies at 1:200 dilution. After 3 washes with PBS, the samples were incubated for 10 min with DAPI (1 µg/mL) in PBS, followed by a final wash in PBS. Fluorescence images were captured at 10× or 20× on a Zeiss LSM 710/780 confocal microscope and processed using ImageJ.
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