10–30 µM PRX-IIE in 100 mM Tris-HCl, pH 8, was reduced for 30 min at room temperature (RT) with 4 mM DTT. Desalting was achieved by passing the solution through PD10 columns. S-glutathionylation was carried out by disulfide exchange with oxidized glutathione (GSSG) for 1 h at RT. Excess GSSG was removed via acetone precipitation. Afterward, S-glutathionylation was detected by Western blot using a monoclonal anti-GSH antibody (Thermo Scientific, Schwerte, Germany). In addition, molecular masses of modified and unmodified proteins were assessed by ESI-MS as mentioned before. For deglutathionylation, 10 µM glutathionylated PRX-IIE was incubated with 10 µM of GRX-S12, GRX-C5, or SRX and 0.5 mM GSH at 25 °C. The decrease of glutathionylated PRX-IIE was determined using Western Blot with anti-GSH antibodies. The spot intensities were analyzed using ImageJ.
Monoclonal anti gsh antibody
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The Monoclonal anti-GSH antibody is a laboratory reagent used for the detection and quantification of glutathione (GSH) in biological samples. It is a highly specific antibody that binds to GSH, enabling researchers to measure GSH levels in cells, tissues, or other biological matrices.
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Lab products found in correlation
2 protocols using monoclonal anti gsh antibody
1
Redox Regulation of Peroxiredoxin II E
2
Probing Redox-Dependent Protein Modifications
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10 -30 µM PRX-IIE in 100 mM Tris-HCl, pH 8, was reduced for 30 min at room temperature (RT) with 4 mM DTT. Desalting was achieved by passing the solution through PD10 columns. Sglutathionylation was carried out by disulfide exchange with oxidized glutathione (GSSG) for 1 h at RT. Excess GSSG was removed via acetone precipitation. Afterwards, S-glutathionylation was detected by Western blot using monoclonal anti-GSH antibody (Thermo Scientific, Schwerte, Germany). In addition, molecular masses of modified and unmodified proteins were assessed by ESI-MS as mentioned before. For deglutathionylation, 10 µM glutathionylated PRX-IIE was incubated with 10 µM of GrxS12, GrxC5 or SRX and 0.5 mM GSH at 25 °C. The decrease of glutathionylated PRX-IIE was determined using Western Blot with anti-GSH antibodies. The spot intensities were analysed using ImageJ.
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