Astragaloside IV and calycosin were purchased from Nanjing Plant Origin Biological., PR China, with a purity of 98%. HMGB1 was purchased from Sino Biological Inc (Beijing, China), with the purity of HMGB1 over 90% determined by SDS-PAGE. The absorption spectra were detected on Cary Series ultraviolet-visible (UV–vis) spectrophotometer (Agilent Technologies, USA). The excitation and emission spectra were scanned on Cary Eclipse Fluorescence Spectrophotometer. The CD spectrum was measured on JASCO 810 spectropolarimeter. Other experimental chemicals were purchased from Sinopharm Chemical Reagent Co., Ltd (Beijing, China).
Hmgb1
Manufactured by Sino Biological
Sourced in China
HMGB1 is a protein that functions as a DNA-binding protein and regulates gene transcription. It is involved in various cellular processes, including DNA repair, inflammation, and cell differentiation. HMGB1 is commonly used in scientific research to study its biological functions and potential applications.
Automatically generated - may contain errors
Lab products found in correlation
810 spectropolarimeter, by Jasco (1 mentions)
Celltracker green, by Thermo Fisher Scientific (1 mentions)
μ slide chamber, by Ibidi (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Fibronectin, by Merck Group (1 mentions)
Tnf α, by Sino Biological (1 mentions)
Formyl methionyl leucyl phenylalanine (fmlp), by Merck Group (1 mentions)
Ifn γ, by Sino Biological (1 mentions)
Interleukin 4 (il 4), by Sino Biological (1 mentions)
Tgf β, by Sino Biological (1 mentions)
Osteo assay surface, by Corning (1 mentions)
Macrophage colony stimulating factor, by Novoprotein (1 mentions)
3 protocols using hmgb1
1
Characterization of Astragaloside IV and Calycosin Binding to HMGB1
2
Neutrophil Extracellular Vesicle Formation Assay
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Neutrophils (5 × 106 cells) were stained cell tracker green (1.5-2.0 μg/mL, Invitrogen) and incubated in either μ-slide chamber (Ibidi) pre-coated with fibronectin (5 μg/mL, Merck Millipore) for evaluation of NDTR formation or in confocal plates for evaluation of NDMV formation. Neutrophils were stimulated with various stimulators: fMLP (1 μM, Sigma-Aldrich), LPS (1 μg/mL, Sigma-Aldrich), C5a (50 ng/mL, Sino Biologicals), S100B (100 ng/mL, Sino Biologicals), HMGB1 (100 ng/mL, Sino Biologicals), TNF-α (50 ng/mL, Sino Biologicals), IFN-γ (100 ng/mL, Sino Biologicals), TGF-β (20 ng/mL, Sino Biologicals), IL-4 (20 ng/mL, Sino Biologicals), PMA (100 μg/mL, Sigma-Aldrich), and N omega-Nitro-L-arginine methyl ester hydrochloride (L-NAME, 10 μM, Tocris bioscience). Then, EV formations were visualized by immunofluorescence microscopy (Olympus IX83, Olympus) at 37 °C, 5% CO2 for 1 hour.
3
Saquinavir Modulates Osteoclastogenesis
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Mouse bone marrow–derived cells were extracted from the femora and tibiae of 4- to 6-week-old female C57BL/6 mice in accordance with the Institutional Animal Care and Use Committee Guide (Tenth People’s Hospital of Tongji University, Shanghai, People’s Republic of China). Briefly, bone marrow was flushed from the medullary cavities and seeded onto a 10-cm-diameter cell culture dish overnight. Unattached cells were collected and reseeded in medium containing α-minimum essential medium, 10% fetal bovine serum, and 1% penicillin-streptomycin at 1 × 104 cells/well on a 96-well plate (Osteo Assay Surface; Corning, Corning, N.Y.) for TRAP staining and 1 × 105 cells/well on a six-well plate to collect RNA samples. After 24 hours, the media were changed to basal medium containing 30 ng/ml macrophage colony-stimulating factor (Novoprotein Scientific, Shanghai, People’s Republic of China) 30 ng/ml for 3 days. Subsequently, cells were cultured in 30 ng/ml macrophage colony-stimulating factor, 100 ng/ml receptor activator of nuclear factor kappa-Β ligand (Novoprotein Scientific), with/without supplementation of various concentrations of saquinavir (0.05, 0.25, 0.5, and 2.5 μg/ml) or 100 ng/ml HMGB1 (Sino Biological, Inc., Beijing, People’s Republic of China) for up to 8 days for TRAP staining and 7 days for quantitative polymerase chain reaction analysis as indicated.
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