DRG were dissociated with a mortar and pestle in tissue extraction reagent supplied with protease and phosphatase inhibitors as previously described.27 (link),28 (link) Assay kits for glutathione (703002, Cayman Chemical, Ann Arbor, MI, USA) (detection range: 0.25–16 μM) superoxide dismutase (706002, Cayman Chemical) (detection range: 0.005–0.050 U/ml), DNA/RNA oxidative damage (589320, Cayman Chemical) (detection range: 10.3–3,000 pg/ml), IL-1β (RLB00, R&D Systems, Minneapolis, MN) (detection range: 5–2,000 pg/ml), TNF (RTA00, R&D Systems) (detection range: 5–800 pg/ml), and CCL2 (ab100778, Abcam) (detection range: 24.67–18,000 μM), were used according to manufacturer instructions. Results were normalized to total protein levels (Bradford protein assay).
Ab100778
Manufactured by Abcam
Sourced in United States
Ab100778 is a laboratory equipment product offered by Abcam. It serves as a core function, but no further details can be provided in a concise, unbiased, and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Ab213906, by Abcam (2 mentions)
Lxsarm, by R&D Systems (2 mentions)
Ab119558, by Abcam (2 mentions)
Rta00, by R&D Systems (1 mentions)
Assay kits for glutathione, by Cayman Chemical (1 mentions)
Tissueruptor, by Qiagen (1 mentions)
Era32rb, by Thermo Fisher Scientific (1 mentions)
Ab105136, by Abcam (1 mentions)
3 protocols using ab100778
1
Oxidative Stress and Inflammatory Markers Assay
2
Inflammatory Cytokine Analysis in Atrial Tissue
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Atrial tissue was minced and homogenized using a tissue homogenizer (TissueRuptor, QIAGEN) according to the manufacturer protocol. Inflammatory cytokines and chemokines were measured using a multiplex Luminex-based assay (LXSARM, R&D Systems). Each sample was run in duplicate in a 96-well plate. IL-1β, IL-2, IL-18, and TNF-α were measured using a MAGPIX system (C4447b, Luminex). Acquired mean fluorescence data were analyzed and calculated by xPONENT software. IL-6 (ERA32RB, Invitrogen), TGF-β1 (ab119558, Abcam), PDGF-AB (ab213906, Abcam), and MCP-1 (ab100778, Abcam) were quantified using commercially available ELISAs according to the manufacturers’ protocols.
3
Atrial Inflammation Biomarkers Analysis
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Homogenized atrial tissue obtained after the invasive electrophysiological study was analyzed for markers of atrial inflammation using ELISAs and a custom multiplex Luminex-based assay (LXSARM, R&D Systems). Interleukin 6 (IL-6, ERA32RB, Life Sciences), IL-17 (MBS2022678, MyBioSource), platelet-derived growth factor AB (PDGF-AB, ab213906, Abcam), monocyte chemoattractant protein-1 (MCP-1, ab100778, Abcam) and transforming growth factor beta 1 (TGFβ1; ab119558, Abcam) were analyzed using commercially available ELISAs. Four analytes (IL-1β, IL-10, IL-18, tumor necrosis factor alpha (TNFα)) were measured using a MAGPIX system (LXSARM-06) with post-hoc mean fluorescence quantification using xPONENT software. Atrial myeloperoxidase activity was evaluated using a colorimetric assay according to the manufacturer's directions (ab105136, Abcam).
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