WB
Western blot analysis of Glutaminase on different lysates with Rabbit anti-Glutaminase antibody at 1/1,000 dilution. Lane 1: 293T cell lysate (20 µg/Lane), Lane 2: HeLa cell lysate (20 µg/Lane), Lane 3: K-562 cell lysate (20 µg/Lane), Lane 4: PC-12 cell lysate (20 µg/Lane), Lane 5: Rat brain tissue lysate (30 µg/Lane), Exposure time: 24 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.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Glutaminase 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 human tonsil tissue with Rabbit anti-Glutaminase 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.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Glutaminase with Rabbit anti-Glutaminase antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Glutaminase antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.IF-P
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Glutaminase 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.FC
Flow cytometric analysis of HeLa cells labeling Glutaminase. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Glutaminase Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Glutamine is an important molecule involved in several cellular functions, including nitrogen and carbon transport, hepatic urea synthesis, renal ammoniagenesis, and gluconeogenesis. Glutamine is catabolized by either the liver-type (LGA) or kidney-type (KGA) glutaminase. KGA is mitochondrial specific protein whose expression in kidney is increased during metabolic acidosis. This process is mediated by an 8-base AU-sequence in KGA that functions as a pH-response element. The human KGA gene maps to chromosome 2, and produces three isoforms, designated KGA, GAC, and GAM, by alternative splicing. KGA is synthesized as a cytosolic protein that is transported to the mitochondria as an intermediate protein, and is further cleaved into the KGA isoform and the GAC isoform. The processing of the GAM isoform is unclear. The KGA isoform is abundant in brain and kidney, while the GAC isoform is principally expressed in cardiac muscle and pancreas. The GAM isoform is solely expressed in cardiac and skeletal muscle. |
| Immunogen | Synthetic peptide within human Glutaminase aa 50-100. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIF-PIHCWB |
| WB:1:1000-1:2000 IHC:1:200-1:1000 ICC/IF:1:50-1:200 IF-P:1:50-1:200 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | AAD20 antibody DKFZp686O15119 antibody FLJ10358 antibody GAC antibody GAM antibody GLS antibody GLS1 antibody GLSK_HUMAN antibody Glutaminase C antibody Glutaminase kidney isoform antibody Glutaminase phosphate activated antibody K-glutaminase antibody KGA antibody KIAA0838 antibody L-glutamine amidohydrolase antibody mitochondrial antibody |
|---|---|
| Molecular Weight(MW) | 74kDa(Observed band size: 65kDa) |
| Cellular Localization | Cytoplasm, Mitochondrion. |
WB
Western blot analysis of Glutaminase on different lysates with Rabbit anti-Glutaminase antibody at 1/1,000 dilution. Lane 1: 293T cell lysate (20 µg/Lane), Lane 2: HeLa cell lysate (20 µg/Lane), Lane 3: K-562 cell lysate (20 µg/Lane), Lane 4: PC-12 cell lysate (20 µg/Lane), Lane 5: Rat brain tissue lysate (30 µg/Lane), Exposure time: 24 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.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Glutaminase 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 human tonsil tissue with Rabbit anti-Glutaminase 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.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Glutaminase with Rabbit anti-Glutaminase antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Glutaminase antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
IF-P
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Glutaminase 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.
FC
Flow cytometric analysis of HeLa cells labeling Glutaminase. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000-1:2000 IHC:1:200-1:1000 ICC/IF:1:50-1:200 IF-P:1:50-1:200 FC:1:1000 |
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| 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. |
