Cells were grown in coated 96-well plates or chamber slides, fixed in 4% paraformaldehyde for 10 min, and permeabilized in 0.3% Triton X-100 in PBS for 10 min. Cells were then blocked with 5% BSA for at least 1 hr before incubation with chicken anti-GFP (Abcam, ab13970) 1:2000, rabbit anti-Tom20 (Santa Cruz, SC11415) 1:500, mouse anti-SSEA4 (Cell Signaling CS 4755) 1:500, mouse anti Tra-1-60 (Cell Signaling CS 4746) 1:500, mouse anti Tra-1-81 (Cell Signaling CS 4745) 1:500, rabbit anti-Pax6 (Covance, PRB-278P) 1:500, or mouse anti-Nestin (Abcam, ab6142) 1:1000 overnight at 4°C. Cells were washed 3 times with PBS, then incubated with AlexaFluor 488, AlexaFluor 555, and/or AlexaFluor 647 (Fisher) at 1:500 dilutions for 1 hr at RT. To stain nuclei, cells were incubated with 1:10000 Hoechst 33342 (Life Technologies). Cells were preserved in Prolong Gold anti-fade reagent (Life Technologies) and imaged on a fluorescence microscope (Z1 Axio Observer Apotome, Zeiss). Mitochondrial length was quantified using an ImageJ mitochondrial morphometry plugin as described previously (Dickey and Strack, 2011 (link)).
Mouse anti tra 1 60
Manufactured by Cell Signaling Technology
Mouse anti Tra-1-60 is a primary antibody that recognizes the Tra-1-60 antigen, which is a specific marker for undifferentiated human embryonic stem cells and induced pluripotent stem cells. This antibody can be used for the identification and characterization of these cell types.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti ssea4, by Cell Signaling Technology (3 mentions)
Ab13970, by Santa Cruz Biotechnology (2 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
Mouse anti nestin, by Abcam (2 mentions)
Rabbit anti tom20, by Santa Cruz Biotechnology (2 mentions)
Mouse anti tra 1 81, by Cell Signaling Technology (2 mentions)
Alexa fluor 555, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 647, by Thermo Fisher Scientific (2 mentions)
Z1 axio observer apotome, by Zeiss (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions)
Rabbit anti pax6, by Fortrea (2 mentions)
Goat anti sox2, by Santa Cruz Biotechnology (1 mentions)
Goat anti oct3 4, by Santa Cruz Biotechnology (1 mentions)
Mouse anti tra 1 81, by Merck Group (1 mentions)
Tcs sp5 laser scanning microscope, by Leica (1 mentions)
Rabbit anti foxa2, by Cell Signaling Technology (1 mentions)
Goat anti pdx1 antibody, by R&D Systems (1 mentions)
3 protocols using mouse anti tra 1 60
1
Immunocytochemistry and Mitochondrial Analysis
2
Immunofluorescence Characterization of Stem Cells
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Cells were fixed with 4% paraformaldehyde for 30 min and then permeabilized in PBS containing 0.2% Triton X-100. Cells were blocked with PBS containing 3% BSA, and incubated with primary antibodies overnight at 4 °C. Then secondary antibodies were incubated for 1 h at room temperature after washing with PBS. Images were acquired on a TCS SP5 laser-scanning microscope (Leica). The following antibodies and dilutions were used: goat anti-OCT-3/4 (1:500, #sc-8628, Santa Cruz), goat anti-SOX2 (1:500, #sc-17320, Santa Cruz), mouse anti-TRA-1-60 (1:1000, #4746, Cell Signaling), mouse anti-TRA-1-81 (1:50, MAB4381, Millipore), mouse anti-SSEA4 (1:500, #4755, Cell Signaling), rabbit anti-FOXA2 (1:250, #8186, Cell Signaling), goat anti-SOX17 antibody (1:500, #GT15094, Acris/Novus), goat anti-PDX1 antibody (1:500, #AF2419, R&D Systems).
3
Immunocytochemistry and Mitochondrial Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were grown in coated 96-well plates or chamber slides, fixed in 4% paraformaldehyde for 10 min, and permeabilized in 0.3% Triton X-100 in PBS for 10 min. Cells were then blocked with 5% BSA for at least 1 hr before incubation with chicken anti-GFP (Abcam, ab13970) 1:2000, rabbit anti-Tom20 (Santa Cruz, SC11415) 1:500, mouse anti-SSEA4 (Cell Signaling CS 4755) 1:500, mouse anti Tra-1-60 (Cell Signaling CS 4746) 1:500, mouse anti Tra-1-81 (Cell Signaling CS 4745) 1:500, rabbit anti-Pax6 (Covance, PRB-278P) 1:500, or mouse anti-Nestin (Abcam, ab6142) 1:1000 overnight at 4°C. Cells were washed 3 times with PBS, then incubated with AlexaFluor 488, AlexaFluor 555, and/or AlexaFluor 647 (Fisher) at 1:500 dilutions for 1 hr at RT. To stain nuclei, cells were incubated with 1:10000 Hoechst 33342 (Life Technologies). Cells were preserved in Prolong Gold anti-fade reagent (Life Technologies) and imaged on a fluorescence microscope (Z1 Axio Observer Apotome, Zeiss). Mitochondrial length was quantified using an ImageJ mitochondrial morphometry plugin as described previously (Dickey and Strack, 2011 (link)).
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