WB
Western blot analysis of GCLM on different lysates with Rabbit anti-GCLM antibody at 1/1,000 dilution. Lane 1: A431 cell lysate, Lane 2: NIH/3T3 cell lysate, Lane 3: PC-12 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 59 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of GCLM on different lysates with Rabbit anti-GCLM antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: Mouse liver tissue lysate (20 µg/Lane) Lane 3: Rat liver tissue lysate (20 µg/Lane) Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-GCLM antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-GCLM antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-GCLM antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IF-F
Immunofluorescence analysis of frozen mouse hippocampus tissue labeling GCLM with Rabbit anti-GCLM antibody. The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/50 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue).IF-F
Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling GCLM with Rabbit anti-GCLM antibody. The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue).| Product Name | GCLM Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Gamma-glutamylcysteine synthetase (γ-GCS) is the rate limiting enzyme for glutathione (L-gamma-glutamyl-L-cysteinylglycine, GSH) synthesis. GSH is ubiquitous in mammalian cells as a vital intra- and extracellular protective antioxidant. γ-GCS is a heterodimer of a heavy catalytic subunit and a light regulatory subunit that is responsive to inflammation, phenolic antioxidants, heat shock, oxidants and cytokines. The human gamma-GCS gene encoding the 367 amino acid catalytic subunit maps to chromosome 6p12. The human γ-GCS gene encoding the regulatory subunit maps to chromosome 1p22-p21. The two subunits of γ-GCS form a heterodimeric zinc metalloprotein that gains activity through formation of a reversible disulfide bond. |
| Immunogen | Recombinant protein within Human GCLM aa 225-274 / 274. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | IF-FIHCWB |
| WB:1:1000-1:2000 IHC:1:200-1:1000 IF-F:1:50 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | Gamma ECS regulatory subunit antibody Gamma-ECS regulatory subunit antibody Gamma-glutamylcysteine synthetase regulatory subunit antibody GCLM antibody GCS light chain antibody GLCLR antibody Glutamate cysteine ligase regulatory subunit antibody Glutamatecysteine ligase modifier subunit antibody Glutamatecysteine ligase regulatory subunit antibody GSC light chain antibody GSH0_HUMAN antibody |
|---|---|
| Molecular Weight(MW) | 31 kDa |
| Cellular Localization | Cytosol. |
WB
Western blot analysis of GCLM on different lysates with Rabbit anti-GCLM antibody at 1/1,000 dilution. Lane 1: A431 cell lysate, Lane 2: NIH/3T3 cell lysate, Lane 3: PC-12 cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 59 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of GCLM on different lysates with Rabbit anti-GCLM antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: Mouse liver tissue lysate (20 µg/Lane) Lane 3: Rat liver tissue lysate (20 µg/Lane) Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-GCLM antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-GCLM antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-GCLM antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IF-F
Immunofluorescence analysis of frozen mouse hippocampus tissue labeling GCLM with Rabbit anti-GCLM antibody. The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/50 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue).
IF-F
Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling GCLM with Rabbit anti-GCLM antibody. The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue).| Application Notes | WB:1:1000-1:2000 IHC:1:200-1:1000 IF-F:1:50 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
