Psb12379
Manufactured by Bio-Techne
Sourced in United Kingdom, United States
The PSB12379 is a laboratory equipment product from Bio-Techne. It is designed for use in scientific research and testing applications. The core function of this product is to provide a specific technical capability, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
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Lab products found in correlation
Wortmannin, by Cell Signaling Technology (2 mentions)
U0126, by Cell Signaling Technology (2 mentions)
Dpcpx, by Bio-Techne (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
N formyl methionyl leucyl phenylalanine, by Merck Group (1 mentions)
Cxcl1, by BioLegend (1 mentions)
8 cyclopentyl 1 3 dipropylxanthine dpcpx, by Merck Group (1 mentions)
Ultra low attachment 6 well plate, by Corning (1 mentions)
Psb1115, by Bio-Techne (1 mentions)
Zm241385, by Bio-Techne (1 mentions)
Theophylline, by Merck Group (1 mentions)
7 protocols using psb12379
1
Purchasing Chemicals for Research
2
Screening Assay for CD39 and CD73 Inhibitors
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Applied chemicals and kits: 8-cyclopentyl-1,3-dipropylxanthine (CPX or DPCPX), 8-cyclopentyl-N3-[3-(4-(fluorosulfonyl)benzoyloxy)propyl]-N1-propylxanthine (FSCPX), sodium polyoxotungstate (POM-1; as a part of the CD39 Inhibitor Screening Assay Kit, see below), disodium N6-benzyl-α,β-methyleneadenosine-5′-diphosphate (PSB-12379), CD39 Inhibitor Screening Assay Kit, and CD73 Inhibitor Screening Assay Kit.
CPX and DMSO were purchased from Merck KGaA (Darmstadt, Germany). FSCPX was manufactured by Santa Cruz Biotechnology, Inc. (Heidelberg, Germany) and distributed by BIO-Kasztel, Ltd. (Budapest, Hungary). Kits (including POM-1) were produced by BPS Bioscience (San Diego, CA, USA) and distributed by THP Medical Products Vertriebs GMBH (Vienna, Austria). PSB-12379 was manufactured by Tocris Bioscience (Bristol, UK) and distributed by Bio-Techne R&D Systems, Ltd. (Budapest, Hungary). Redistilled water was used to dissolve and dilute POM-1 as well as dilute the pre-dissolved PSB-12379. FSCPX and CPX were dissolved in dimethyl-sulfoxide (DMSO; purchased from Merck KGaA (Darmstadt, Germany)), and then, they were diluted with redistilled water (when needed).
CPX and DMSO were purchased from Merck KGaA (Darmstadt, Germany). FSCPX was manufactured by Santa Cruz Biotechnology, Inc. (Heidelberg, Germany) and distributed by BIO-Kasztel, Ltd. (Budapest, Hungary). Kits (including POM-1) were produced by BPS Bioscience (San Diego, CA, USA) and distributed by THP Medical Products Vertriebs GMBH (Vienna, Austria). PSB-12379 was manufactured by Tocris Bioscience (Bristol, UK) and distributed by Bio-Techne R&D Systems, Ltd. (Budapest, Hungary). Redistilled water was used to dissolve and dilute POM-1 as well as dilute the pre-dissolved PSB-12379. FSCPX and CPX were dissolved in dimethyl-sulfoxide (DMSO; purchased from Merck KGaA (Darmstadt, Germany)), and then, they were diluted with redistilled water (when needed).
3
Inhibition and Activation of Purinergic Pathways
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The selective CD39 inhibitor sodium polyoxotungstate (POM1), CD73 inhibitor PSB 12,379 (N6-Benzyl-α,β-methyleneadenosine 5’-diphosphate disodium salt), and adenosine receptor agonist 1-(6-Amino-9 H-purin-9-yl)-1-deoxy-N-ethyl-β-D-ribofuranuronamide (NECA) were from Tocris (Bristol, UK). The CD39 mimic potato apyrase was from Sigma, and recombinant human (rh)CD73 was from Daresbury Proteins (Warrington, UK).
4
Neutrophil and Monocyte Migration Assay
5
Evaluating CD73 Inhibition in 2D and 3D Noz Cell Cultures
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Adherent and spheroid NOZ cells were seeded in 96-well plates and were exposed for 72 h to PSB 12379 (CD73 pharmacological inhibitor) (Tocris) at indicated doses. The MTS reagent was then added to each well and absorbance was measured at 490 nm. Enumeration of spheroids was performed by initially seeding spheroids at 2000 cells/well in a 6-well ultra-low-attachment plates (Corning Incorporated). Spheroids were cultured with serum-free stem cell-conditioned medium. PSB 12379 was added at a concentration of 10 µM to the spheroid culture. Number of spheres per culture well were counted on day 7 using an inverted microscope equipped with a digital camera (Olympus DP21).
6
Modulation of Macrophage Polarization by MSC Exosomes
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To investigate the modulatory effects of MSC exosomes on macrophage polarization, the rat primary macrophages were treated with either 10 μg/mL exosomes or PBS vehicle for 24 and 48 h (Figure 1 A). To investigate the role of CD73 in mediating the exosome effects, macrophages were co-treated with 10 μg/mL exosomes and 10 nM PSB12379 (CD73 inhibitor; Tocris Bioscience, Bristol, UK) (Figure 1 B). The role of adenosine receptors in the activation of AKT and ERK pathways was investigated by pre-treating the cells with 2 μM of ZM241385 (A2A receptor antagonist; Tocris), 200 nM PSB1115 (A2B receptor antagonist; Tocris), 1 μM wortmannin (AKT inhibitor; Cell Signaling Technology, Danvers, MA, USA), or 10 μM U0126 (ERK inhibitor; Cell Signaling Technology) for 1 h, before treatment with 10 μg/mL exosomes (Figure 1 C). For the inhibitor studies, the macrophages were harvested at 30 min for western blot analysis, 24 and 48 h for gene expression analysis, and 48 h for immunofluorescence staining (Figure 1 B,C). All in vitro experiments were performed in triplicates (n = 3) in two independent trials.
7
Exosome-Mediated Odontogenic Differentiation
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P3 DPCs were seeded at a density of 2 × 104 cells/cm2 and cultured for 16 h before the medium was changed to low serum culture medium for 24 h, and then treated with 1, 5, 10 μg/ml exosomes or PBS for over 48 h. To investigate the involvement of adenosine receptors and activation of AKT and ERK pathways, cells were pre-treated with 1 mM theophylline (a non-selective inhibitor of adenosine receptors) (Sigma) [34] (link), 1 µM wortmannin (AKT inhibitor) or 10 µM U0126 (ERK inhibitor) (Cell Signaling Technology, Danvers, MA, USA) or equivalent volume of distilled water or dimethyl sulfoxide (DMSO) as vehicle controls for 1 h, before treatment with 10 μg/ml exosomes or PBS for 48 h. In experiment with CD73 inhibitor, N6-Benzyl-α,β-methyleneadenosine 5′-diphosphate disodium salt (PSB12379; Tocris Biosciences, CO, USA), cells were co-treated with exosomes and 10 nM PSB12379 for 1 h. Following treatment, cells were rinsed with PBS and harvested for analysis. In odontogenic differentiation experiments, cells were treated with 10 µg/ml exosomes or PBS in odontogenic medium with or without the inhibitors. Medium change with fresh exosomes and inhibitors was performed every alternate day till day 7.
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