Dissociated single cell suspensions from primary tumors (as described previously (24 (link))) or cultured cells were incubated with Aldefluor reagent (Stem Cell Technologies) for 30 minutes according to manufacturer’s instructions. Cells were then rinsed before incubation with CD61-biotin for 20 minutes at 4°C. Following that, cells were rinsed and incubated with other antibodies marking CSCs for 20 minutes at 4°C. Cells were then rinsed and resuspended in Aldefluor buffer before sorting or analysis by FACSAria or FACSCanto instruments (BD Biosciences). Flow cytometry data were analyzed using FlowJo software.
Protocol detail
Identification of Cancer Stem Cells
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : University of Cincinnati Medical Center
Top 5 similar protocols
Protocol cited in 4 other protocols
Variable analysis
independent variables
- Incubation with Aldefluor reagent for 30 minutes
- Incubation with CD61-biotin for 20 minutes at 4°C
- Incubation with other antibodies marking CSCs for 20 minutes at 4°C
- Cell sorting or analysis by FACSAria or FACSCanto instruments (BD Biosciences)
- Flow cytometry data analyzed using FlowJo software
- Dissociated single cell suspensions from primary tumors or cultured cells
- Rinsing of cells before incubation with antibodies
- Resuspension of cells in Aldefluor buffer before sorting or analysis
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!