TROMA-1, a rat monoclonal antibody against cytokeratin Endo A, a marker of primitive endoderm, and a mouse monoclonal antibody (mAb MC-480) that recognizes the stem cell-specific cell surface antigen (SSEA-1), were purchased from the Developmental Studies Hybridoma Bank (University of Iowa, IA, USA). All trans-retinoic acid (RA) was purchased from Sigma (USA) and was dissolved in ethanol (100 mM stock). Puromycin was purchased from Sigma (USA). The plasmid vector pCDNA3, Neomycin (G418) were purchased from Invitrogen (USA). Plasmid harboring non-targeting short hairpin RNA (shRNA), pLKO.1 control vector was purchased from Open Biosystems, USA.
Mc 480
Manufactured by Developmental Studies Hybridoma Bank
Sourced in United States
The MC-480 is a laboratory instrument designed for the cultivation and maintenance of hybridoma cell lines. It features a temperature-controlled incubation chamber and programmable settings to ensure optimal growth conditions for the cells.
Automatically generated - may contain errors
Lab products found in correlation
Neomycin g418, by Thermo Fisher Scientific (1 mentions)
Trans retinoic acid, by Merck Group (1 mentions)
Puromycin, by Merck Group (1 mentions)
Troma 1, by Developmental Studies Hybridoma Bank (1 mentions)
Cyan analyzer, by Beckman Coulter (1 mentions)
Live dead fixable dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
2 protocols using mc 480
1
Stem Cell Surface Antigen Identification
2
SSEA-1 Expression Analysis by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were trypsinized, washed once in PBS, and resuspended in PBS; dead cells were stained using the LIVE/DEAD Fixable Dead Cell Stain Kit (Invitrogen) according to the manufacturer’s instructions. Next, cells were fixed in 2% paraformaldehyde (PFA) for 15 min at room temperature, washed with PBS and stained with anti-SSEA-1 mouse monoclonal IgM antibody (mc-480, University of Iowa, Developmental Studies Hybridoma Bank, 1:400) at 37°C for 20 min. After a PBS wash, the cells were labeled with fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgM (Millipore, 1:200) for 15 min at 37°C. After one PBS wash, stained cells were resuspended in PBS containing 1% FBS and 3% BSA, and at least 10,000 events of healthy cells were analyzed with a CyAN Analyzer (Beckman Coulter) and FlowJo ver.10 software (Tree Star, Ashland, USA). Control cells were not treated with primary antibody.
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!