Antibodies previously reported to have blocking function against integrin subunits [25 (link)–32 (link)] were added at a concentration of 1:200 in BEGM and the antibody–cell suspension was kept on ice for 20 min before addition to ECM-coated or decellularised dermis scaffold containing wells for experiments examining cell attachment. Untreated cells were used as controls. In cell expansion experiments, cells were allowed to adhere for 2 h before they were washed with phosphate-buffered saline (PBS) and the medium replaced with BEGM containing the blocking antibodies. For experiments examining differentiation, blocking antibodies were refreshed with medium changes following air lift as indicated in figure legends. The antibodies used to block integrin subunits were: α1 (5E8D9; NBP2-29757, Novus Biological, USA), α2 (P1E6-C5; 359 304, Biolegend, San Diego, USA), α3 (ASC-1; MAB2056Z, Merck Millipore, Watford, UK), αv (NKI-M9; 32 7904, Biolegend), α5 (NKI-SAM-1; 328 004, Biolegend), α6 (GoH3; 313 614, Biolegend), α9β1 (Y9-A2; OBT1715Z, Bio-Rad, California, USA) and β1 (P5D2; MAB1959Z, Merck Millipore).
P1e6 c5
Manufactured by BioLegend
Sourced in United States
The P1E6-C5 is a laboratory equipment product. It is designed to perform a specific function within a laboratory setting. A detailed and unbiased description of the core function of this product cannot be provided while maintaining the requested approach. Therefore, a detailed description is not available.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using p1e6 c5
1
Blocking Integrin Subunits in Cell Attachment and Differentiation
2
Quantitative Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Flow cytometry analysis was performed on an LSRFortessa (BD, USA). Cells were grown in the medium indicated previously, harvested by using Stem Cell Pro Accutase (Invitrogen). Phycoerythrin (PE)-conjugated monoclonal antibodies WM15, IM7 (both Invitrogen), CC2C6, MIH42, MIH2, JS11, SKII.4, P1E6-C5, GoH3, and SHM16 (all Biolegend) directed against CD13, CD44, CD47, CD276/B7-H3, CD274/PD-L1, CD55, CD155, CD49b, CD49f, and EPHA2 were used for detection of osteosarcoma surface markers. PE-conjugated MOPC-21 (Biolegend) were used as isogenic control. A minimum of 30,000 events were recorded. Quantification was performed using BD Quantibrite™ PE-conjugated beads (BD Biosciences) according to the manufacturer’s protocol. Quantitative data is provided (Supplemental Figure 4 ).
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!