To assess the changes in implantation protein profiles in BBT-EVs as a result of BBT-MSC in vitro communication, pre-confluent 100 mm2 tissue culture dishes of BBT-9 and BBT-18 were cultured for 48 h in media supplemented with MSC-EVs isolated by SEC from stimulated or unstimulated MSC. Confluent 100 mm2 tissue culture dishes of eMSC or pbMSC were cultured for 48 h with or without IFN-τ stimulation (MBS1131960, Mybiosource) (1000 ng/mL). Then, EVs from the different eMSC or pbMSC lines were pooled separately. In addition, to assess uterine cytokines effect on BBT-EVs, pre-confluent 100 mm2 tissue culture dishes of BBT-9 and BBT-18 were cultured in media supplemented with Activin A (PHC9564, Thermo Fisher Scientific) (100 ng/mL), Activin A, and FSTL1 (10924H08H50, Thermo Fisher Scientific) (100 and 50 ng/mL, respectively) or without cytokines during 48 h. The experimental design is shown in Figure 5 . Bead-assisted flow cytometry was performed on the BBT-EVs obtained following the procedures described to analyze the implantation marker expression of MMP2 (MA1-772, Thermo Fisher Scientific); HSPH1 (PA5-77793, Thermo Fisher Scientific); TDGF1 (NB100-1597, Novus Biologicals), and PEG3 (TA343590, Acris-antibodies).
Nb100 1597
Manufactured by Novus Biologicals
NB100-1597 is a laboratory instrument used for detecting and quantifying proteins in biological samples. It is a multi-mode microplate reader capable of various detection modes, including absorbance, fluorescence, and luminescence. This product is suitable for a wide range of applications in life science research and development.
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2 protocols using nb100 1597
1
Implantation Protein Profiling in BBT-EVs
2
Exploring Implantation Markers in BBT-EVs
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To assess the changes in implantation protein pro les in BBT-EVs as a result of BBT-MSC in vitro communication, pre-con uent 100-mm 2 tissue culture dishes of BBT-9 and BBT-18 were cultured for 48h in media supplemented with MSC-EVs isolated by SEC from stimulated or unstimulated MSC. Con uent 100-mm 2 tissue culture dishes of eMSC or pbMSC were cultured for 48 h with or without IFN-τ stimulation (MBS1131960, Mybiosource) (1000 ng/ml). Then, EVs from the different eMSC or pbMSC lines were pooled separately. In addition, to assess uterine cytokines effect on BBT-EVs, pre-con uent 100mm 2 tissue culture dishes of BBT-9 and BBT-18 were cultured in media supplemented with Activin A (PHC9564, Thermo Fisher Scienti c) (100 ng/ml), Activin A and FSTL1 (10924H08H50, Thermo Fisher Scienti c) (100 and 50 ng/ml respectively) or without cytokines during 48h. The experimental design is shown in Fig. 1. Bead-assisted ow cytometry was performed to BBT-EVs obtained following the procedures besides described to analyze the implantation marker expression of MMP2 (MA1-772, Thermo Fisher Scienti c); HSPH1 (PA5-77793, Thermo Fisher Scienti c); TDGF1 (NB100-1597, Novus Biologicals) and PEG3 (TA343590, Acris-antibodies).
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