Glutathione reductase Antibody [E19C20]

Application: Reactivity: Human, Mouse, Rat

Lane 1: Neuro-2a, Lane 2: IMR-32, Lane 3: SH-SY5Y, Lane 4: JEG-3

Usage Information

Dilution
WB1:100-1:1000 IHC1:50-1:500 IF1:50-1:500

Application
WB, IP, IHC, IF, ELISA

Reactivity
Human, Mouse, Rat

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
50-65 kDa

Positive Control	Human fallopian tube tissue; Human parathyroid gland tissue; IMR-32 cells; JEG-3 cells; C6 cells; EOC 20 cells; H4 cells; SH-SY5Y cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
Glutathione reductase Antibody [E19C20] detects endogenous levels of total Glutathione reductase protein.

Clone
E19C20

Synonym(s)
GSR; Glutathione reductase, mitochondrial; EC:1.8.1.7; GR; GRase; GLUR; GRD1

Background
Glutathione reductase (GSR) is a dimeric flavoprotein enzyme belonging to the pyridine nucleotide-disulfide oxidoreductase family, essential for maintaining cellular redox homeostasis by catalyzing the NADPH-dependent reduction of oxidized glutathione (GSSG) to two molecules of reduced glutathione (GSH). GSH serves as the primary non-protein thiol antioxidant, detoxifying reactive oxygen species, peroxides via glutathione peroxidases, and xenobiotics through conjugation. Each monomer of glutathione reductase has an N-terminal FAD-binding Rossmann fold that holds the isoalloxazine ring, a central NADPH-binding domain with precise hydride-transfer geometry to position the nicotinamide ring close to the FAD, and a C-terminal dimerization/interface domain that forms the GSSG-binding pocket at the subunit interface. The enzyme possesses a redox-active disulfide (Cys58–Cys63 in humans) adjacent to FAD, where NADPH reduces FAD to FADH⁻ through a hydride transfer, generating a nucleophilic Cys58-thiolate that attacks the proximal sulfur of GSSG to form a mixed disulfide and release the first GSH. His467 then protonates the departing thiolate, and an intramolecular thiol-disulfide exchange with Cys63 regenerates the active disulfide, liberating the second GSH. This ping-pong bi-bi mechanism ensures high catalytic turnover and maintains a high GSH to GSSG ratio, which is crucial for defending against oxidative stress in the cytosol and mitochondria.

Background

Glutathione reductase (GSR) is a dimeric flavoprotein enzyme belonging to the pyridine nucleotide-disulfide oxidoreductase family, essential for maintaining cellular redox homeostasis by catalyzing the NADPH-dependent reduction of oxidized glutathione (GSSG) to two molecules of reduced glutathione (GSH). GSH serves as the primary non-protein thiol antioxidant, detoxifying reactive oxygen species, peroxides via glutathione peroxidases, and xenobiotics through conjugation. Each monomer of glutathione reductase has an N-terminal FAD-binding Rossmann fold that holds the isoalloxazine ring, a central NADPH-binding domain with precise hydride-transfer geometry to position the nicotinamide ring close to the FAD, and a C-terminal dimerization/interface domain that forms the GSSG-binding pocket at the subunit interface. The enzyme possesses a redox-active disulfide (Cys58–Cys63 in humans) adjacent to FAD, where NADPH reduces FAD to FADH⁻ through a hydride transfer, generating a nucleophilic Cys58-thiolate that attacks the proximal sulfur of GSSG to form a mixed disulfide and release the first GSH. His467 then protonates the departing thiolate, and an intramolecular thiol-disulfide exchange with Cys63 regenerates the active disulfide, liberating the second GSH. This ping-pong bi-bi mechanism ensures high catalytic turnover and maintains a high GSH to GSSG ratio, which is crucial for defending against oxidative stress in the cytosol and mitochondria.

References
https://pubmed.ncbi.nlm.nih.gov/30605126/ https://pubmed.ncbi.nlm.nih.gov/2584829/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%