The cells were mechanically dissociated to obtain a single-cell suspension in a solution containing 2 mM EDTA (Sigma-Aldrich) and 0.5% FBS (Sigma-Aldrich) in PBS. The cells were harvested by centrifugation (500 × g, 5 min, 4°C) and incubated with a PE-conjugated anti-CD133 antibody (#85-12-1339-73, eBioscience) for 2 h on ice. They were then incubated or 2 h on ice with secondary antibody conjugated to phycoerythrin, washed with PBS twice, filtered through a 70-µm cup filter (Falcon; Becton Dickinson), and kept on ice until FACS analysis. COS-7 cells were used as a negative control, and liver tumor cells with high CD133 expression (kindly provided by Dr. Jun Li, Hepatological Surgery Department, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei Province, China) were used as a positive control. Cell sorting was performed using a FACS Vantage TS flow cytometer (Becton Dickinson).
Pe conjugated anti cd133 antibody
Manufactured by Thermo Fisher Scientific
The PE-conjugated anti-CD133 antibody is a laboratory reagent used for the detection and analysis of CD133-expressing cells. CD133 is a cell surface marker commonly used to identify and isolate stem and progenitor cells. The PE-conjugation allows for the fluorescent labeling and visualization of CD133-positive cells using flow cytometry or other compatible analytical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Bodipy 493 503, by Thermo Fisher Scientific (1 mentions)
Facs influx instrument, by BD (1 mentions)
Penicillin streptomycin, by Beyotime (1 mentions)
Rpmi 1640, by Corning (1 mentions)
2 protocols using pe conjugated anti cd133 antibody
1
Isolation and Sorting of CD133+ Cells
2
Multiparametric Analysis of PDO Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells from PDOs were prepared using 300 units/mL collagenase IV (Worthington, LS004210) at 37°C for 30 minutes, followed by three 5-minute washes with RPMI-1640 (Corning) supplemented with 10% FBS (MilliporeSigma) and 50 units/mL penicillin/streptomycin (Beyotime). Cells were treated with BD Fix Buffer I and Perm Buffer III and were stained for 45 minutes at room temperature with the following antibodies: PE-conjugated anti-CD133 antibody (eBiosciences, 12-1338-42), anti–human OPA1 antibody (Cell Signaling Technology, 67589), and anti–human SPDEF antibody (Santa Cruz Biotechnology Inc., sc-166846). BODIPY 493/503 (Thermo Fisher Scientific, D3922) was used to quantify the intracellular accumulation of lipid droplets. Cells were analyzed using BD Canto II FACS analysis, and for some experiments, live cells were sorted with the BD FACS Influx instrument. Fluorescence minus one (FMO) was used for the gating strategy. Data were analyzed with FlowJo software.
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!