Anti human cd253 trail antibody

In order to confirm the presence of TRAIL on the membrane of the genetically modified MSCs 1 × 10⁵ native MSCs, MSCs-TRAIL and MSCs-BFP were detached from the culture flask using Accutase™ cell dissociation reagent (#A1110501, Gibco, Waltham, MA, USA) and washed once with PBS. The washed cells were stained with anti-human CD253 (TRAIL) antibody (#308210, BioLegend, San Diego, CA, USA) for 30 min at room temperature in the dark. The stained cells were washed with PBS and analyzed by flow cytometry using FACSAria III (BD Biosciences, San Jose, CA, USA).

In order to evaluate the presence of the TRAIL gene mRNA in the CIMVs isolated from native MSCs (native CIMVs), MSCs-BFP (CIMVs-BFP), and MSCs-TRAIL (CIMVs-TRAIL) the tRNA was extracted from the newly isolated CIMVs, and qPCR was performed as described in Section 2.4.
In order to evaluate the presence of the TRAIL protein, the native CIMVs, CIMVs-BFP, and CIMVs-TRAIL were isolated, as previously described, and lysed in an RIPA buffer containing a Halt™ Protease and Phosphatase Inhibitor Cocktail (Thermo Scientific, Waltham, MA, USA). Then, 50 µg of each CIMV was used for the following Western blot analysis, which was performed as described in Section 2.5.
In order to analyze the localization of the TRAIL protein, the native CIMVs, CIMVs-BFP, and CIMVs-TRAIL were isolated, as previously described, and stained with anti-human CD253 (TRAIL) antibody (#308210, BioLegend, San Diego, CA, USA) for 30 min at room temperature in the dark. The staining was assessed using the FACSAria III flow cytometer (BD Biosciences, San Jose, CA, USA).