The cells were rinsed briefly with phosphate-buffered saline (PBS) and fixed for 20 min in 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4) at room temperature. The cells were permeabilized for 10 min with 0.1% Triton X-100 in PBS, and blocked for 45–60 min with 4% bovine serum albumin in PBS at room temperature. Cells were incubated overnight at 4 °C with one of the following antibodies: anti-Oct4 (1:500; Abcam, Cambridge, MA, USA), anti-Sox2 (1:500; NB110-37235, Novus Biologicals, Littleton, CO, USA), anti-Nanog (1:500; Abcam, Cambridge, MA, USA), anti-c-Myc (1:250; bs-4963R, Bioss, Woburn, MA, USA), anti-Klf4 (1:250, bs-1064R, Bioss, Woburn, MA, USA). This was followed by incubation with the following secondary antibody: Alexa Fluor 488-labeled anti-rabbit IgG (1:500; Invitrogen, Carlsbad, CA, USA). Nuclei were counterstained using DAPI (1 mg/mL PBS; Invitrogen, Carlsbad, CA, USA).
Nb110 37235
Manufactured by Novus Biologicals
Sourced in United States
The NB110-37235 is a laboratory instrument designed for cell culture applications. It functions as an incubator, providing a controlled environment for the growth and maintenance of cell lines. The product specifications and technical details are available upon request.
Automatically generated - may contain errors
Lab products found in correlation
Anti mouse alexa fluor 488, by Thermo Fisher Scientific (4 mentions)
Oct 3 4 sc 5279, by Santa Cruz Biotechnology (2 mentions)
Anti rabbit, by Thermo Fisher Scientific (2 mentions)
Facsaria, by BD (2 mentions)
Alexa fluor 488 labeled anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Anti nanog, by Abcam (1 mentions)
Anti oct4, by Abcam (1 mentions)
Bs 4963r, by Bioss Antibodies (1 mentions)
Anti sox2, by Novus Biologicals (1 mentions)
Anti c myc, by Bioss Antibodies (1 mentions)
Anti klf4, by Bioss Antibodies (1 mentions)
Bs 1064r, by Bioss Antibodies (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
3 protocols using nb110 37235
1
Immunofluorescence Staining of Stem Cell Markers
2
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 were harvested using 0.5 mM EDTA in PBS, washed with PBS, and then fixed with ice-cold 70% ethanol. The fixed cells were washed twice with wash buffer (5mM EDTA, 2% Foetal Bovine Serum in PBS) and incubated for 1 hour with the primary antibodies SOX2 (NB110-37235, Novus Biological, 1:400) or Oct-3/4 (sc-5279, Santa Cruz, 1:400). The cells were then washed and incubated with secondary antibodies anti-Rabbit or anti-Mouse AlexaFluor 488 (Thermo Fisher Scientific, 1:1000) for 1 hour. For TRA-1-60 expression assessment, non-fixed cells and the TRA-1-60 antibody (MAB4360, Millipore, 1:100) were used. Flow cytometry analysis was performed using BD FACSAria™ (BD Biosciences). Datasets were analysed using FlowJo software (www.flowjo.com).
3
Characterization of Pluripotent Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested using 0.5 mM EDTA in PBS, washed with PBS, and then fixed with ice-cold 70% ethanol. The fixed cells were washed twice with wash buffer (5mM EDTA, 2% Foetal Bovine Serum in PBS) and incubated for 1 hour with the primary antibodies SOX2 (NB110-37235, Novus Biological, 1:400) or Oct-3/4 (sc-5279, Santa Cruz, 1:400). The cells were then washed and incubated with secondary antibodies anti-Rabbit or anti-Mouse AlexaFluor 488 (Thermo Fisher Scientific, 1:1000) for 1 hour. For TRA-1-60 expression assessment, non-fixed cells and the TRA-1-60 antibody (MAB4360, Millipore, 1:100) were used. Flow cytometry analysis was performed using BD FACSAria™ (BD Biosciences). Datasets were analysed using FlowJo software (www.flowjo.com).
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