GSK4112, SR8278, SR1001, SR1078, lithium chloride and staurosporine (STS) were purchased from Tocris. Drugs were dissolved in DMSO or water and stored as concentrated solutions at −80°C prior to use.
Sr1001
Manufactured by Bio-Techne
Sourced in United Kingdom
SR1001 is a laboratory reagent product manufactured by Bio-Techne. It is a small molecule compound used for research purposes. The core function of SR1001 is to act as a selective agonist for a specific nuclear receptor.
Automatically generated - may contain errors
Lab products found in correlation
Sr1078, by Bio-Techne (2 mentions)
Staurosporine, by Bio-Techne (1 mentions)
Sr8278, by Bio-Techne (1 mentions)
Gsk4112, by Bio-Techne (1 mentions)
Dimethyl sulfoxide (dmso), by Bio-Techne (1 mentions)
Pen step, by Merck Group (1 mentions)
Complete dmem, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Biosera (1 mentions)
3 protocols using sr1001
1
Drug Compound Preparation Protocol
2
Preparation of RORα/γ Agonist and Antagonist
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RORα/γ agonist (SR1078; mw 431.25) and antagonist (SR1001; mw 477.4) were both purchased from TocrisBioscience (Avonmouth, Bristol, United Kingdom). Stock solutions of 10 mM were prepared by diluting 10 mg of SR1078 and SR1001 in 2.3188 ml and 2.0947 ml, respectively, of sterile filtered DMSO (TocrisBioscience, Avonmouth, Bristol, United Kingdom). The stock of each was stored at -20 °C as light-protected 50 µL aliquots. Desired concentrations were calculated by dividing the initial stock concentration by the desired concentrations to get the dilution factor.
3
Assessing RORγt Expression in MAIT Cells
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Human dermal fibroblasts at passage 20 were cultured in complete DMEM (Sigma) supplemented with 1% pen/step (Sigma) and 10% FBS (BioSera). At 80% confluency, cells were treated with recombinant IL-17 (20ng/ml) for 24 hours before harvesting for mRNA extraction and RT-PCR analysis as previously described.
MAIT cell RORgt analysis. MAIT cell RORgt expression in patients and controls was determined by transcription factor flow cytometry. PBMC, CD45+ skin cells or expanded MAIT cells were activated (TCR dynabeads) for 18 hours before determining RORgt expression using a specific RORgt mAb (BD Biosciences) and transcription factor staining kit (Biolegend). To assess the effect of RORgt inhibition on IL-17 production the specific inhibitor SR1001 (Tocris) was added at a final concentration of 5 μM to purified MAIT cells from either healthy donors or patients with H.S.
MAIT cell RORgt analysis. MAIT cell RORgt expression in patients and controls was determined by transcription factor flow cytometry. PBMC, CD45+ skin cells or expanded MAIT cells were activated (TCR dynabeads) for 18 hours before determining RORgt expression using a specific RORgt mAb (BD Biosciences) and transcription factor staining kit (Biolegend). To assess the effect of RORgt inhibition on IL-17 production the specific inhibitor SR1001 (Tocris) was added at a final concentration of 5 μM to purified MAIT cells from either healthy donors or patients with H.S.
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