Glutaminase Recombinant Rabbit Monoclonal Antibody [SN68-09]
cat.: ET1611-5
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Rat, Mouse
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SN68-09
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 74 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Glutaminase aa 50-100.
Positive control: 293T cell lysate, HeLa cell lysate, K-562 cell lysate, PC-12 cell lysate, rat brain tissue lysate, HeLa, HepG2, human tonsil tissue, human kidney tissue.
Subcellular location: Cytoplasm, Mitochondrion.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
Uniprot #: SwissProt: O94925 Human | D3Z7P3 Mouse | P13264 Rat
Alternative names: AAD20 DKFZp686O15119 FLJ10358 GAC GAM GLS GLS1 GLSK_HUMAN Glutaminase C Glutaminase kidney isoform Glutaminase phosphate activated K-glutaminase KGA KIAA0838 L-glutamine amidohydrolase mitochondrial
Images
ET1611-5_1.jpg Fig1: Western blot analysis of Glutaminase on different lysates with Rabbit anti-Glutaminase antibody (ET1611-5) 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)

Predicted band size: 74 kDa
Observed band size: 65 kDa

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 (ET1611-5) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1611-5_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling Glutaminase with Rabbit anti-Glutaminase antibody (ET1611-5) 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 (ET1611-5) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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.
ET1611-5_3.jpg Fig3: ICC staining of Glutaminase in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1611-5_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Glutaminase antibody (ET1611-5) 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 (ET1611-5) 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.
ET1611-5_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Glutaminase antibody (ET1611-5) 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 (ET1611-5) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.