Product nameAnti-GLUD1 antibody
See all GLUD1 primary antibodies
DescriptionRabbit polyclonal to GLUD1
SpecificityDue to high sequence homology with GLUD2 there is a potential for cross-reactivity.
Tested applicationsSuitable for: Dot blot, WB, ELISA, ICC/IF, IP, IHC-Pmore details
Species reactivityReacts with: Mouse, Rat, Cow, Human, Pig
Full length native protein (purified) corresponding to Cow GLUD1. Bovine liver
- This antibody gave a positive result in IHC in the following FFPE tissue: Human normal liver. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: MCF-7
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
Concentration information loading...
Purification notesThis antibody was purified from monospecific antiserum by delipidation and defibrination.
Our Abpromise guarantee covers the use of ab34786 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Dot blot||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 62 kDa.|
|ELISA||1/4000 - 1/16000.|
|ICC/IF||1/1000. PubMed: 20925087|
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionMay be involved in learning and memory reactions by increasing the turnover of the excitatory neurotransmitter glutamate.
Involvement in diseaseDefects in GLUD1 are the cause of familial hyperinsulinemic hypoglycemia type 6 (HHF6) [MIM:606762]; also known as hyperinsulinism-hyperammonemia syndrome (HHS). Familial hyperinsulinemic hypoglycemia [MIM:256450], also referred to as congenital hyperinsulinism, nesidioblastosis, or persistent hyperinsulinemic hypoglycemia of infancy (PPHI), is the most common cause of persistent hypoglycemia in infancy and is due to defective negative feedback regulation of insulin secretion by low glucose levels. In HHF6 elevated oxidation rate of glutamate to alpha-ketoglutarate stimulates insulin secretion in the pancreatic beta cells, while they impair detoxification of ammonium in the liver.
Sequence similaritiesBelongs to the Glu/Leu/Phe/Val dehydrogenases family.
Cellular localizationMitochondrion matrix.
- Information by UniProt
- AI118167 antibody
- DHE3_HUMAN antibody
- EC 126.96.36.199 antibody
Glutamate Dehydrogenase 1 was immunoprecipitated using 0.5mg Mouse Liver tissue lysate, 5µg of Rabbit polyclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Mouse Liver tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab34786.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 62kDa, non specific bands - 30 and 82kDa: We are unsure as to the identity of this extra band; Glutamate Dehydrogenase
ICC/IF image of ab34786 stained MCF-7 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 ab34786 at 1/1000 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.
IHC image of GLUD1 staining in Human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab34786, 1µg/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.
This product has been referenced in:
- Wang T et al. L-leucine stimulates glutamate dehydrogenase activity and glutamate synthesis by regulating mTORC1/SIRT4 pathway in pig liver. Anim Nutr 4:329-337 (2018). Read more (PubMed: 30175263) »
- Andersen JV et al. Impaired Hippocampal Glutamate and Glutamine Metabolism in the db/db Mouse Model of Type 2 Diabetes Mellitus. Neural Plast 2017:2107084 (2017). WB ; Mouse . Read more (PubMed: 28695014) »