Evaluation of the expression of mesenchymal stem cell surface markers (CD73 and CD44), embryonic stem cell surface marker (OCT-4), and hematopoietic cell marker (CD45) was done by flow cytometric analysis as described previously [28 (link)]. Cells (105 cells/100 μl) were gently washed in PBS containing 2% FBS and incubated separately with PE-conjugated mouse anti-human CD73 (561014; BD Pharmingen), CD44 (550989; BD Pharmingen), and CD45 (560975; BD Pharmingen) in darkness for at least 40 min at 4 °C. Evaluation of OCT-4 expression was analysed using indirect intracellular flow cytometry. The cells were permeabilized with 0.1% saponin. Then primary rabbit antihuman OCT-4 antibody (ab19857, Abcam) was added for 40 min and then incubated with FITC-conjugated goat anti-rabbit Ig (Sigma-Aldrich) for 30 min. As negative controls, isotype IgG (555748; BD Pharmingen) was used. Afterward, cells were washed twice with PBS-FBS, fixed in 1% formaldehyde solution, and analyzed by a flow cytometer (Partec GmbH).
Fitc conjugated goat anti rabbit ig
Manufactured by Merck Group
FITC-conjugated goat anti-rabbit Ig is a laboratory reagent used to detect and visualize rabbit immunoglobulins (Ig) in various immunoassays and imaging techniques. The product consists of goat-derived antibodies specific to rabbit Ig molecules that have been conjugated with the fluorescent dye fluorescein isothiocyanate (FITC).
Automatically generated - may contain errors
2 protocols using fitc conjugated goat anti rabbit ig
1
Mesenchymal and Hematopoietic Cell Markers Analysis
2
Immunophenotyping of Mesenchymal Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As mentioned by Darzi S, et al30 (link), aliquots of 105 cells/100 μl were incubated separately with phycoerythrin (PE)-conjugated mouse anti human CD29 (clone 04-MAR; BD Pharmingen), CD73 (clone AD2; BD Pharmingen), CD44 (clone 515; BD Pharmingen), CD133 (clone TMP4; eBioscience), and CD105 (clone 43A3; BioLegend) for 40 min at 4°C.
To assess Octamer-binding transcription factor 4 (OCT-4) expression, the cells were washed with 0.1% saponin-permeabilized cells and treated with primary rabbit antihuman OCT-4 antibody (Abcam) for 40 min and then incubated with FITC-conjugated goat anti-rabbit Ig (Sigma-Aldrich) for 30 min. Isotype IgG was used as negative controls (clone MOPC-21; BD Pharmingen). Afterwards, all cell suspensions were washed twice with PBS-FBS, fixed in 1% formaldehyde solution, and analyzed using a flow cytometer (Partec GmbH, Munster, Germany) and appropriate isotype controls.
To assess Octamer-binding transcription factor 4 (OCT-4) expression, the cells were washed with 0.1% saponin-permeabilized cells and treated with primary rabbit antihuman OCT-4 antibody (Abcam) for 40 min and then incubated with FITC-conjugated goat anti-rabbit Ig (Sigma-Aldrich) for 30 min. Isotype IgG was used as negative controls (clone MOPC-21; BD Pharmingen). Afterwards, all cell suspensions were washed twice with PBS-FBS, fixed in 1% formaldehyde solution, and analyzed using a flow cytometer (Partec GmbH, Munster, Germany) and appropriate isotype controls.
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!