antibody

Anti-Glutathione Reductase/GSR Picoband Antibody

A01479-1

100 ug/vial

polyclonal

domestic rabbit

nonconjugated

human, mouse, rat

western blot, ELISA, immunocytochemistry, flow cytometry, immunohistochemistry - paraffin section, immunohistochemistry - frozen section

Enabled

Anti-Glutathione Reductase/GSR Picoband Antibody

Primary Antibodies, Polyclonal Antibodies, IHC ICC IF Antibodies

GSR

Unconjugated / $280 APC / $330 APC-Cy7 / $330 FITC / $330 PE / $330 PE-Cy5 / $330 PE-Cy7 / $330 100ug / $280 100ug+Free HRP Secondary BA1054 / $280 100ug+Free Biotin Secondary BA1003 / $280

Polyclonal

0.5-1mg/ml, actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.

No

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na 2 HPO 4 , 0.05mg NaN 3 .

Rabbit IgG polyclonal antibody for Glutathione Reductas detection. Tested with WB, IHC-P, IHC-F, ICC/IF, FCM, Direct ELISA in Human;Mouse;Rat.

Rabbit IgG polyclonal antibody for Glutathione Reductas detection. Tested with WB, IHC-P, IHC-F, ICC/IF, FCM, Direct ELISA in Human;Mouse;Rat.

100 ug/vial

P00390

Rabbit

E. coli-derived human Glutathione Reductase recombinant protein (Position: K256-R522).

Lyophilized

At -20 C for one year. After reconstitution, at 4 C for one month. It can also be aliquotted and stored frozen at -20 C for a longer time. Avoid repeated freezing and thawing.

No cross reactivity with other proteins.

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Western blot, 0.1-0.5ug/ml. Immunohistochemistry(Paraffin-embedded Section), 0.5-1ug/ml. Immunohistochemistry(Frozen Section), 0.5-1ug/ml. Immunocytochemistry/Immunofluorescence, 2ug/ml. Flow Cytometry, 1-3ug/1x10 6 cells. Direct ELISA, 0.1-0.5ug/ml.

ELISA, Flow Cytometry, IF, IHC-P, IHC-F, ICC, WB

Human, Mouse, Rat

Figure 1. Western blot analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, . Lane 2: human HepG2 whole cell lysates, . Lane 3: human A549 whole cell lysates, . Lane 4: human 22RV1 whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Glutathione Reductase antigen affinity purified polyclonal antibody (Catalog # A01479-1) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Glutathione Reductase at approximately 55KD. The expected band size for Glutathione Reductase is at 55KD. Figure 2. Western blot analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat spleen tissue lysates,. Lane 2: rat lung tissue lysates,. Lane 3: rat liver tissue lysates,. Lane 4: rat kidney tissue lysates,. Lane 5: mouse spleen tissue lysates,. Lane 6: mouse lung tissue lysates,. Lane 7: mouse liver tissue lysates,. Lane 8: mouse kidney tissue lysates,. Lane 9: mouse testis tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Glutathione Reductase antigen affinity purified polyclonal antibody (Catalog # A01479-1) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Glutathione Reductase at approximately 55KD. The expected band size for Glutathione Reductase is at 55KD. Figure 3. IHC analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Glutathione Reductase was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-Glutathione Reductase Antibody (A01479-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen. Figure 4. IHC analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Glutathione Reductase was detected in paraffin-embedded section of mouse small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-Glutathione Reductase Antibody (A01479-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen. Figure 5. IHC analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Glutathione Reductase was detected in paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-Glutathione Reductase Antibody (A01479-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen. Figure 6. Flow Cytometry analysis of U20S cells using anti-GSR antibody (A01479-1). Overlay histogram showing U20S cells stained with A01479-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GSR Antibody (A01479-1,1ug/1x10 6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10ug/1x10 6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x10 6 ) used under the same conditions. Unlabelled sample (Red line) was also used as a control. Figure 7. Flow Cytometry analysis of A549 cells using anti-GSR antibody (A01479-1). Overlay histogram showing A549 cells stained with A01479-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-GSR Antibody (A01479-1,1ug/1x10 6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10ug/1x10 6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x10 6 ) used under the same conditions. Unlabelled sample (Red line) was also used as a control. Figure 8. IHC analysis of Glutathione Reductase using anti-Glutathione Reductase antibody (A01479-1). Glutathione Reductase was detected in immunocytochemical section of A549 cell. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti- Glutathione Reductase Antibody (A01479-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen. Figure 9. IF analysis of Glutathione Reductase using anti- Glutathione Reductase antibody (A01479-1) . Glutathione Reductase was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/mL rabbit anti- Glutathione Reductase Antibody (A01479-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. Figure 10. Flow Cytometry analysis of A431 cells using anti- Glutathione Reductase antibody (A01479-1). Overlay histogram showing A431 cells stained with A01479-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Glutathione Reductase Antibody (A01479-1,1ug/1x10 6 cells) for 30 min at 20°C. DyLight?488 conjugated goat anti-rabbit IgG (BA1127, 5-10ug/1x10 6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x10 6 ) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Glutathione reductase (GR), also known as glutathione-disulfide reductase (GSR), is an enzyme that in humans is encoded by the GSR gene. This gene encodes a member of the class-I pyridine nucleotide-disulfide oxidoreductase family. This enzyme is a homodimeric flavoprotein. It is a central enzyme of cellular antioxidant defense, and reduces oxidized glutathione disulfide (GSSG) to the sulfhydryl form GSH, which is an important cellular antioxidant. Rare mutations in this gene result in hereditary glutathione reductase deficiency. Multiple alternatively spliced transcript variants encoding different isoforms have been found.

Cancer, Cell Biology, Metabolism, Mitochondrial, Mitochondrial Markers, Mitochondrial Metabolism, Oxidative Stress, Pathways And Processes, Redox Metabolism, Signal Transduction

Glutathione reductase, mitochondrial; GR; GRase; GSR; GLUR; GRD1

glutathione-disulfide reductase

Maintains high levels of reduced glutathione in the cytosol.

Isoform Mitochondrial: Mitochondrion.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.

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