Protein Expression Analysis in Samples

Total proteins were extracted from SM, resolved on SDS-PAGE gels and transferred to nitrocellulose membranes in a semi-dry Trans-Blot device (Bio-Rad, Madrid, Spain) as described (24 (link), 28 (link)). The following primary antibodies were applied overnight at 4°C: anti-human COX-2 (Santa Cruz Biotechnology, Dallas TX, USA), anti-rabbit IL-6, anti-rabbit TNF, anti-rabbit IL-10, anti-rabbit TGF-β (all from Cloud-Clone Corp; 1/250 dilution); anti-human NLRP3 (AdipoGen, Liestal, Switzerland; 1/1000 dilution); anti-human Caspase-1 (Thermo Fisher Scientific, IL, USA; 1/500 dilution), anti-rabbit IL-18 and IL-1β antibodies (Cloud-Clone Corp, Houston TX, USA; 1/250 dilution). Protein loading control was performed employing EZBlue gel staining reagent (Sigma-Aldrich (11 (link), 24 (link)).

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Medina J.P., Bermejo-Álvarez I., Pérez-Baos S., Yáñez R., Fernández-García M., García-Olmo D., Mediero A., Herrero-Beaumont G, & Largo R. (2023). MSC therapy ameliorates experimental gouty arthritis hinting an early COX-2 induction. Frontiers in Immunology, 14, 1193179.

Publication 2023

Trans-Blot anti-human COX-2 anti-rabbit IL-6 anti-rabbit TNF EZBlue gel staining reagent

Antibodies Caspase 1 Clone Cox 2 Device Dilution Human Il 10 Il 18 Il 1β Membranes Nitrocellulose Protein Rabbit Sds page Tgf β

Corresponding Organization : Universidad Autónoma de Madrid

Other organizations : Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas, Centre for Biomedical Network Research on Rare Diseases

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Variable analysis

independent variables

Total proteins extracted from SM
Primary antibodies applied overnight at 4°C

dependent variables

Protein expression levels of COX-2, IL-6, TNF, IL-10, TGF-β, NLRP3, Caspase-1, IL-18, and IL-1β

control variables

SDS-PAGE gels for protein separation
Nitrocellulose membranes for protein transfer
Semi-dry Trans-Blot device (Bio-Rad, Madrid, Spain) for protein transfer
EZBlue gel staining reagent (Sigma-Aldrich) for protein loading control

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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