The fluorescence reaction of the sulphate diester in VSMCs was tested using 7‐azido‐4 methylcoumarin (C‐7Az, Sigma), which has been shown to selectively respond to H2S.10 (link) VSMCs were incubated with 50 μmol/L C‐7Az PBS for 30 minutes and then washed with PBS. The fluorescence response of C‐7Az in VSMCs was detected using a fluorescence microscope (Olympus, XSZ‐D2) after excitation with a 720 nm laser. The results indicated that excitation fluorescence imaging was useful for detecting H2S by triggering the fluorescence reaction of C‐7Az.
C 7az
Manufactured by Merck Group
Sourced in United States
C-7Az is a piece of laboratory equipment designed for use in scientific research and analysis. It is a compact, versatile instrument that can perform a variety of laboratory tasks. The core function of C-7Az is to provide accurate and reliable measurements and data collection for researchers and scientists. However, a more detailed description is not available while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Xsz d2, by Olympus (6 mentions)
C 7az, by Olympus (3 mentions)
Dcfh da, by Thermo Fisher Scientific (1 mentions)
H2s donor nahs, by Merck Group (1 mentions)
Keap 1 sirna, by Santa Cruz Biotechnology (1 mentions)
Mitosox, by Thermo Fisher Scientific (1 mentions)
Mitotempo, by Merck Group (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
Palmitate, by Merck Group (1 mentions)
9 protocols using c 7az
1
Detecting H2S in Vascular Smooth Muscle Cells
2
Fluorescence Imaging of H2S in RAECs
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The fluorescence intensity of H2S in RAECs was tested using 7‐azido‐4‐methylcoumarin (C‐7Az, Sigma‐Aldrich, USA), which has been proved to selectively respond to H2S. RAECs were incubated with 50 μM C‐7Az PBS for 30 min., followed by washing of the cells with PBS for three times. Visualization of the turn‐on fluorescence response of C‐7Az to H2S in RAECs was carried out using a fluorescence microscope (Olympus, XSZ‐D2, Japan). These results confirmed that excitation fluorescence imaging could be used to detect H2S through the triggered fluorescence response of C‐7Az.
3
Fluorescence Imaging of H2S in RAECs
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The turn-on fluorescence response of H2S in RAECs was tested using 7-Azido-4-Methylcoumarin, (C-7Az, Sigma), which has been proved to selectively respond to H2S. RAECs were incubated with 50 μM C-7Az PBS for 30 min, followed by washing of the cells with PBS. Visualization of the turn-on fluorescence response of C-7Az to H2S in RAECs was carried out using fluorescence microscopy with the excitation of a 720 nm laser. These results confirmed that excitation fluorescence imaging could be used to detect H2S through the triggered fluorescence response of C-7Az.
4
H2S Detection in Cardiac Tissue
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The fluorescence response of H2S in cardiac tissue and neonatal rat cardiomyocytes (NRCMs) was detected using the H2S-specific detection probe C-7Az (7-azido-4-methylcoumarin, Sigma-Aldrich). Frozen sections of db/db cardiac tissues or NRCMs were incubated with H2S-specific detection probe C-7Az (50 μmol/L) in phosphate-buffered saline (PBS) for 30 min at 37 °C. These were washed three times with PBS. Fluorescence microscopy (Olympus, XSZ-D2, Tokyo, Japan) was used to detect the fluorescence responses of H2S in the samples.
5
Mitochondrial Dysfunction Regulation
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H2S donor NaHS, palmitate, MDC, CSE inhibitor PPG, autophagy inhibitor 3MA, disulphide bond inhibitor DTT, mitochondrial ROS inhibitor Mito-TEMPO, ROS inhibitor NAC, E1 inhibitor PYR41 and C-7Az were purchased from Sigma-Aldrich (Sigma, St. Louis, MO, USA). Mito-SOX, DCFH-DA and JC-1 were purchased from Invitrogen (Grand Island, NY, USA). SOD and CAT activity assay kits were purchased from Jiancheng Institute of Bioengineering, Nanjing, China. Keap-1 siRNA was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies to cleaved caspase 3 (25546-1-AP), Bax (50599-2-lg), Bcl2 (12789-1-AP), SOD (10269-1-AP), CAT (21260-1-AP), Beclin1 (11306-1-AP), ATG7 (10088-2-AP), P62 (18420-1-AP), LC3 A/B (66139-1-lg), LC3 B (18725-1-AP), Keap-1 (10503-2-AP), ubiquitin (10201-2-AP), VDAC1 (55259-1-AP), GAPDH (10494-1-AP) and β-actin (66009-1-AP) were purchased from Proteintech (Rosemont, IL, USA).
6
Quantifying H2S in Cardiomyocytes
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The H2S content of HL-1 cardiomyocytes was tested with a selective H2S probe, 7-azido-4-methylcoumarin (C-7Az, Sigma-Aldrich). Cells were incubated with PBS containing 50 µM C-7Az at 37 °C in dark for 30 min and washed twice by PBS. The fluorescence was observed using the fluorescence microscope (Olympus XSZ-D2, Japan).
7
Detecting H2S Fluorescence in Mast Cells
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The fluorescence response of H2S in MCs was detected using 7-Azido-4-Methylcoumarin (C-7Az; Sigma-Aldrich; Merck KGaA), as described previously (24 (link),25 (link)). MCs were incubated with 1 ml PBS (containing 12 µl C-7Az) for 30 min at room temperature and then the cells were washed with PBS. The fluorescence activation response of C-7Az to H2S in MCs were visualized using a microscope (Olympus Corporation) and six fields (magnification, ×200) were randomly selected for statistical analysis.
8
Fluorescence Detection of H2S in Cardiac Cells
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The fluorescence response of H2S in cardiac tissues and neonatal rat cardiomyocytes (NRCMs) was tested with 7-azido-4-methylcoumarin (C-7Az, Sigma-Aldrich). This probe has been testified to selectively respond to H2S [16 (link)]. The samples were incubated with C-7Az (50 μmol/L) in phosphate-buffered saline (PBS) at 37 °C for 30 min and then washed three times with PBS. The fluorescence responses in the samples were observed with a fluorescence microscope (Olympus, XSZ-D2, Japan).
9
Quantifying Cellular Hydrogen Sulfide
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Hydrogen sulfide content in cells was assessed using the 7‐azido‐4‐methylcoumarin probe (C7Az, Sigma) under 720 nm laser excitation, observed via a fluorescence microscope (Olympus, XSZD2). Quantification of hydrogen sulfide (H2S) in cardiac tissue and serum employed the dedicated H2S content Kit (Solarbio).
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