Mature HiNs were fixed overnight in 4% PFA, permeabilized with 1% Triton-X-100 for 5 min at room temperature, and then blocked using 6% normal goat serum at room temperature on a platform rocker for 30 min. HiNs were then incubated in primary antibody (hyperphosphorylated tau: AT8 antibody (ThermoFisher (MN1020) 1:1000), TRA-160 (Abcam (ab16288) 1:200), TRA-181 (Abcam (ab16289) 1:200) overnight at 4 °C on a platform rocker. Neurons were then rinsed 3x in 1xPBS, and incubated in secondary antibody (Alexafluor 594, ThermoFisher 1:1000) at room temperature on a platform rocker for two hours. Fluorescent images were then collected using a 20× air objective using a Nikon Eclipse TE 2000-S microscope (Melville, NY, USA) with an XCite series 120 PC illumination source, captured with Nikon Elements software (AR package, Melville, NY, USA). Data are presented as mean fluorescent intensity in randomly selected ROI in each representative image.
Ab16289
Manufactured by Abcam
Ab16289 is a monoclonal antibody that recognizes the protein of interest X. It is suitable for use in various laboratory applications, including immunoassays and immunohistochemistry. The antibody is produced in mice and purified using affinity chromatography.
Automatically generated - may contain errors
Lab products found in correlation
Ab16288, by Abcam (2 mentions)
Mn1020, by Abcam (1 mentions)
Eclipse te2000 s microscope, by Nikon (1 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Ab16287, by Abcam (1 mentions)
Ab106465, by Abcam (1 mentions)
Ab97959, by Abcam (1 mentions)
Sw3015, by Solarbio (1 mentions)
Ab 59545, by Abcam (1 mentions)
Goat anti rabbit mouse igg h l, by Abcam (1 mentions)
P1110, by Solarbio (1 mentions)
2 protocols using ab16289
1
Immunofluorescence Analysis of Neuronal Markers
2
Immunofluorescence Characterization of Pluripotent Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells with sufficient growth were inoculated into a 12-well plate coated with human recombinant laminin. After adhering to the surface of the well, cells were fixed with 4% paraformaldehyde (P1110, Solarbio) for 15 min, blocked with 3% hydrogen peroxide for 15 min, and incubated with 5% bovine serum albumin (BSA) (Solarbio, SW3015) for 15 min. After rinsing with phosphatebuffered saline (PBS), the cells were incubated overnight with primary antibodies at 4°C, followed by incubation with a secondary antibody (goat anti-rabbit/mouse IgG H&L, Abcam) at 37°C for 1 h. The following primary antibodies were used: rabbit anti-Oct-4 (ab59545, Abcam), rabbit anti-Nanog (ab106465, Abcam), rabbit anti-Sox2 (ab97959, Abcam), mouse anti-SSEA-4 (ab16287, Abcam), mouse anti-TRA-1-60 (ab16288, Abcam), and mouse anti-TRA-1-81 (ab16289, Abcam). Subsequently, cell nuclei were stained with 4 0 ,6-diamidino-2-phenylindole (DAPI) for microscopic observation and imaging.
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