Single-cell suspension was prepared in ice-cold PBS containing 5% FBS, and 106 cells were analyzed with 1 μg antibody using FACSCantoII (BD Biosciences) flow cytometer. FITC-conjugated Sca-1 (BD Biosciences, Franklin Lakes, NJ), PE-conjugated Cd44 (BD Biosciences), and APC/Cy7-conjugated Cd45 (Bioregend, San Diego, CA), were used as indicated. FACSDiva software (BD Biosciences) and FlowJo (Tree Star) were used for data acquisition and analysis. For the cell cycle analysis, 106 cells were fixed in 1 ml PBS with 50% ethanol on ice for 1 h. Cells were resuspended in 0.25 ml of PBS with 50 μg RNase (DNase free) and incubated at 37 °C for 1 h. After incubation with 10 μg/ml propidium iodide on ice in dark, cell cycle analysis was performed using a single-parameter histogram with linear x-axis to represent DNA content.
Fitc conjugated sca 1
Manufactured by BD
Sourced in United States
FITC-conjugated Sca-1 is a fluorescent-labeled antibody that binds to the Sca-1 (Stem Cell Antigen-1) protein. Sca-1 is a cell surface marker commonly used to identify and isolate murine hematopoietic stem and progenitor cells. The FITC (Fluorescein Isothiocyanate) conjugation allows for the detection and analysis of Sca-1-positive cells using flow cytometry or fluorescence-based techniques.
Automatically generated - may contain errors
2 protocols using fitc conjugated sca 1
1
Multiparametric Flow Cytometry Analysis
2
Isolation and Quantification of Circulating EPCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Circulating EPCs were isolated according to the methods of our previously published papers [11, 25] . Peripheral blood about 0.7 ml was harvested from anesthetized mice and then kept in cool heparin anticoagulant tube. Mononuclear cells were isolated from blood with Histopaque 1083 (Sigma, St. Louis, MO, USA) density gradient centrifugation (3000 rpm) for 25 minutes. After washing and suspension, samples were stained with FITC-conjugated Sca-1 (2 μg/10 6 cells, BD, San Jose, CA, USA) and PE-conjugated Flk-1 (2 μg/10 6 cells, BD, San Jose, CA, USA) for 1 hour at 4°C. Quantification of Sca-1/Flk-1 doublepositive cells was performed with a BD FACSCalibur Flow cytometer. A non-stained sample was used to set up a threshold, and the isotype specific conjugated anti-IgG was used as a negative control [11, 25] .
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!