Glutamate Dehydrogenase Antibody - BSA Free

Western Blot: Glutamate Dehydrogenase Antibody [NB600-853] - Western blot analysis is shown using Glutamate Dehydrogenase antibody to detect the enzyme from bovine liver preparations. Comparison to a molecular weight marker indicates a predominant band of ~62 kDa. The higher molecular weight band may represent a subunit dimer. A 4-20% gradient gel was used to separate proteins prior to transfer to 0.2 um nitrocellulose. The blot was incubated with a 1:1,000 dilution of the antibody for 2 h at room temperature followed by detection using IRDye(TM)800 labeled Goat-a-Rabbit IgG [H&L] diluted 1:5,000 for 45 min at room temperature. IRDye(TM)800 fluorescence image was captured using the Odyssey(R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.

N/A: Glutamate Dehydrogenase Antibody [NB600-853]

• Reactivity: Hu, Bv
• Applications: WB, ELISA, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

Glutamate Dehydrogenase Antibody - BSA Free Summary

• Description: This product was prepared from monospecific antiserum by a delipidation and defibrination. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-rabbit serum, purified and partially purified Glutamate Dehydrogenase [Bovine Liver]
  
  Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
• Immunogen: This Glutamate Dehydrogenase Antibody was prepared from whole rabbit serum produced by repeated immunizations with a full length Glutamate Dehydrogenase protein isolated from Bovine Liver. (Uniprot: P00366)
• Localization: Mitochondrial matrix
• Marker: Mitochondria Marker
• Isotype: Serum
• Clonality: Polyclonal
• Host: Rabbit
• Gene: GLUD1
• Purity: Delipidation and Defibrination

Applications/Dilutions

• Dilutions:
  • ELISA 1:4000-1:16000
  • Immunohistochemistry
  • Immunoprecipitation 1:100
  • Western Blot 1:1000-1:3000
• Application Notes: This antibody has been tested by western blot. Specific conditions for reactivity should be optimized by the end user. Bovine glutamate dehydrogenase exists as a homohexamer located within the mitochondrial matrix. Expect a band approximately 56 kDa in size corresponding to glutamate dehydrogenase monomer subunit by western blotting in the appropriate cell or tissue extract. Anti-Glutamate Dehydrogenase Antibody is suitable for use in ELISA.
  
  Use in Immunohistochemistry reported in scientific literature (PMID 27623078).

Reactivity Notes

BLAST analysis was used to determine that cross reactivity is suggested for both mitochondrial and brain isoforms (GDH1 and GDH2), from both bovine and human sources. Additionally similar reactivity is suggested for most primate species including green monkey, white gibbon, chimpanzee orangutan, and gorilla. A high degree of sequence homology is also noted for Glutamate Dehydrogenase from chicken, mouse, rat, dog, and other mammals as well as Xenopus tropicalis, zebrafish, rainbow trout and Atlantic salmon.
Human reactivity reported in scientific literature (PMID: 27623078).

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Delipidation and Defibrination

Alternate Names for Glutamate Dehydrogenase Antibody - BSA Free

• EC 1.4.1
• EC 1.4.1.3
• GDH 1
• GDH
• GDH1
• GLUD
• glutamate dehydrogenase (NAD(P)+)
• Glutamate Dehydrogenase 1
• glutamate dehydrogenase 1, mitochondrial
• MGC132003

Background

One enzyme central to the metabolism of glutamate is glutamate dehydrogenase (GDH1; EC 1.4.1.3), that catalyzes the reversible deamination of L-glutamate to 2-oxoglutarate using NAD+ or NADP+. Mammalian GDH is composed of six identical subunits, and the regulation of GDH is very complex. It has been a major goal to identify the substrate and regulatory binding sites of GDH. It is only in recent years that the three-dimensional structure of GDH from microorganisms is available. Very recently, crystallization of bovine liver GDH was reported for the first time from the mammalian sources. However, remarkably little is known about the detailed structure of mammalian GDH, especially the brain enzymes.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Glutamate Dehydrogenase Antibody (NB600-853)(4)

Publications using NB600-853

• Talari NK, Mattam U, Meher NK et al. Lipid-droplet associated mitochondria promote fatty-acid oxidation through a distinct bioenergetic pattern in male Wistar rats Nature communications 2023-02-11 [PMID: 36765117] (WB, Rat)
• Knudsen ES, Balaji U, Freinkman E et al. Unique metabolic features of pancreatic cancer stroma: relevance to the tumor compartment, prognosis, and invasive potential. Oncotarget. 2016-11-29 [PMID: 27623078] (IF/IHC, Human)
• Yang C, Ko B, Hensley CT et al. Glutamine Oxidation Maintains the TCA Cycle and Cell Survival during Impaired Mitochondrial Pyruvate Transport. Mol Cell. 2014-10-21 [PMID: 25458842] (WB, Human)
  
  Details:
  Glutamate Dehydrogenase antibody used for WB of whole-cell lysates and preparations of isolated mitochondria (Mito) or cytosol from SFxL cells - SFxL cells generated by infecting SF188 glioma cells with a retoviral vectorto express human Bcl-xL pBabe-Hygro-Bcl-xL (Figure 2A). Antibody also used on SFxL cells expressing a control shRNA or either of two shRNAs directed against GLUD1 - shGLUD1-A and shGLUD1-B ( Figure 6C )
• Bao X, Pal R, Hascup KN et al. Transgenic expression of Glud1 (glutamate dehydrogenase 1) in neurons: in vivo model of enhanced glutamate release, altered synaptic plasticity, and selective neuronal vulnerability. J Neurosci;29(44):13929-44. 2009-11-04 [PMID: 19890003]

Review for Glutamate Dehydrogenase Antibody (NB600-853) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Pig.

reviewed by: Paula Chen WB Pig 01/09/2019

Summary

• Application: Western Blot
• Sample Tested: Porcine embryo
• Species: Pig
• Lot: 16721

Comments

• Comments: A 4-20% gradient gel was used to separate proteins from pools of 40 blastocysts before transfer to PVDF membranes. The membrane was incubated in 1:1000 anti-glutamate dehydrogenase primary overnight at 4 degrees Celsius. The next day, membranes were incubated in 1:10000 anti-rabbit IgG HRP secondary for 1 hour.

Bioinformatics

• Gene Symbol: GLUD1
• Entrez:
  • 281785(Bovine)
• Uniprot:
  • P00366(Bovine)