Synthetic peptide corresponding to Human Glutaminase aa 516-545 (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH).
WB: 293, 239T and HeLa cell lysates, mouse liver and brain tissue lysates, human brain and rat kidney tissue lysates.
ICC/IF: HeLa and HepG2 cells.
IHC-P: Human liver and mouse brain tissues.
Flow Cyt: HepG2 cells.
Shipped at 4°C. Store at 4°C (up to 6 months). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Preservative: 0.09% Sodium azide Constituent: PBS
Concentration information loading...
Immunogen affinity purified
This antibody is purified through a protein A column, followed by peptide affinity purification.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10 - 1/50.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
1/1000. Predicted molecular weight: 65 kDa.
Use a concentration of 5 µg/ml.
Use at an assay dependent concentration.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Catalyzes the first reaction in the primary pathway for the renal catabolism of glutamine.
KGA is expressed predominantly in brain and kidney but not in liver, isoform 3 is expressed principally in cardiac muscle and pancreas but not in liver or brain, and isoform 2 is expressed solely in cardiac and skeletal muscle.
Belongs to the glutaminase family. Contains 1 ANK repeat.
IHC image of ab93434 staining in human liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab93434, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab93434 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab93434, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse brain tissue labelling Glutaminase with ab93434. A peroxidase-conjugated anti-rabbit IgG was used as the secondary antibody.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Glutaminase (green) with ab93434. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG was used as the secondary antibody. Counter stained with DAPI (blue).
Western blot - Anti-Glutaminase antibody (ab93434)
Anti-Glutaminase antibody (ab93434) at 1/1000 dilution + mouse liver tissue lysate at 35 µg
Secondary Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/5000 dilution
Predicted band size: 65 kDa
Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.
Jiang S et al. Let-7 Suppresses B Cell Activation through Restricting the Availability of Necessary Nutrients. Cell Metab27:393-403.e4 (2018).
Read more (PubMed: 29337138) »
Andersen JV et al. Impaired Hippocampal Glutamate and Glutamine Metabolism in the db/db Mouse Model of Type 2 Diabetes Mellitus. Neural Plast2017:2107084 (2017).
Read more (PubMed: 28695014) »