Intestines from adult Foxm1-hDTR mice were cut open and washed in PBS. Intestinal epithelial cells were separated from mesenchymal cells as described in Fluorescent activated cell sorting and RNA isolation. The crypt epithelium was scraped off and collected. To ensure isolation of pure epithelium, epithelial cells were incubated in 0.05% Trypsin to obtain single cells, followed by incubation with an Alexa 488 mouse EpCAM (Biolegend 1:250) antibody, and sorted in a BD Influx instrument (BD Biosciences). The remaining mesenchymal tissue was washed to remove any epithelial contamination. From the epithelial and mesenchymal fractions, RNA was isolated and cDNA synthesized. For mesenchymal and epithelial-specific markers as well as hDTR, RT-PCR was performed using GoTaq (Promega). Primers used in this experiment are listed in Table 1 .
Alexa 488 mouse epcam
Manufactured by BioLegend
Alexa 488 mouse EpCAM is a fluorescently-labeled antibody that targets the Epithelial Cell Adhesion Molecule (EpCAM) expressed on the surface of epithelial cells. The antibody is conjugated with Alexa Fluor 488, a green fluorescent dye, allowing for the detection and analysis of EpCAM-positive cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using alexa 488 mouse epcam
1
Isolation and Characterization of Intestinal Cell Fractions
2
Isolation and Characterization of Intestinal Epithelial and Mesenchymal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intestines from adult Foxm1–hDTR mice were cut open and washed in PBS. Intestinal epithelial cells were separated from mesenchymal cells as described in fluorescent-activated cell sorting and RNA isolation. The crypt epithelium was scraped off and collected. To ensure isolation of pure epithelium, epithelial cells were incubated in 0.05% trypsin to obtain single cells, followed by incubation with an Alexa-488 mouse EpCAM (1:250; Biolegend, San Diego, CA) antibody, and sorted in a BD Influx instrument (BD Biosciences). The remaining mesenchymal tissue was washed to remove any epithelial contamination. From the epithelial and mesenchymal fractions, RNA was isolated and complementary DNA (cDNA) was synthesized. For mesenchymal and epithelial-specific markers as well as hDTR, reverse-transcription PCR was performed using GoTaq (Promega, Madison, WI). The primers used in this experiment are listed in Table 1 .
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!