Human Dihydrolipoamide Dehydrogenase/DLD Antibody

Catalog #: MAB8646 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB8646-100
MAB8646-SP

Dihydrolipoamide Dehydrogenase/DLD in HeLa Human Cell Line.

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Human Dihydrolipoamide Dehydrogenase/DLD Antibody Summary

Species Reactivity

Human

Specificity

Detects human Dihydrolipoamide Dehydrogenase/DLD in direct ELISAs.

Source

Monoclonal Mouse IgG_2B Clone # 1016619

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived human Dihydrolipoamide Dehydrogenase/DLD
Ala36-Phe509
Accession # P09622

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Immunohistochemistry

5-25 µg/mL

See below

Immunocytochemistry

5-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunocytochemistry

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Dihydrolipoamide Dehydrogenase/DLD in HeLa Human Cell Line. Dihydrolipoamide Dehydrogenase/DLD was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human Dihydrolipoamide Dehydrogenase/DLD Monoclonal Antibody (Catalog # MAB8646) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry

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Dihydrolipoamide Dehydrogenase/DLD in Human Liver Tissue. Dihydrolipoamide Dehydrogenase/DLD was detected in immersion fixed paraffin-embedded sections of human liver tissue using Mouse Anti-Human Dihydrolipoamide Dehydrogenase/DLD Monoclonal Antibody (Catalog # MAB8646) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei in hepatocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Dihydrolipoamide Dehydrogenase/DLD

Dihydrolipoamide dehydrogenase (DLD), also known as LADH, is an NAD-dependent oxidoreductase in the mitochondrial matrix (1). DLD serves as the E3 subunit of four mitochondrial enzyme complexes: pyruvate dehydrogenase, alpha-ketoglutarate dehydrogenase, branched chain alpha-ketotacid dehydrogenase, and the glycine cleavage system (2, 3). It is active as a 120 kDa dimer that catalyzes oxidation within these enzyme complexes. Several mutations of human DLD have been described, some of which contribute to the loss of respiratory function during oxidative stress (4, 5). DLD mutations located at the interface between dimer subunits can impair dimer formation (5, 6). The involvement of DLD in the regulation of lipid peroxidation and lactate metabolism is important for mouse hippocampal neuroblast proliferation and hamster sperm capacitation, respectively (7, 8). DLD polymorphisms in insects can increase their resistance to the pesticide phosphine gas, while they can increase the sensitivity to arsenite in the nematode C. elegans (9). Mature human DLD shares 95-96% amino acid (aa) sequence identity with hamster, mouse, and rat DLD. Alternative splicing generates additional human DLD isoforms that lack the N-terminal 99 aa or carry a deletion of aa 147-194.

References

Brand, M.D. (2010) Exp. Gerontol. 45:466.
Johnson, M.T. et al. (1997) Proc. Natl. Acad. Sci. USA 94:14512.
Sundquist, A.R. and R.C. Fahey (1988) J. Bacteriol. 170:3459.
Ambrus, A. et al. (2011) Hum. Mol. Genet. 20:2984.
Vaubel, R.A. et al. (2011) J. Biol. Chem. 286:40232.
Babady, N.E. et al. (2007) Proc. Natl. Acad. Sci. USA 104:6158.
Calingasan, N.Y. et al. (2008) Neuroscience 153:986.
Panneerdoss, S. et al. (2012) J. Androl. 33:699.
Schlipalius, D.I. et al. (2012) Science 338:807.

Entrez Gene IDs

1738 (Human); 13382 (Mouse); 298942 (Rat)

Alternate Names

2-oxo-glutarate complex, branched chain keto acid dehydrogenase complex); branched chain keto acid dehydrogenase complex; Diaphorase; dihydrolipoamide dehydrogenase (E3 component of pyruvate dehydrogenase complex; Dihydrolipoamide Dehydrogenase; dihydrolipoamide dehydrogenasemitochondrial; DLD; DLDD; DLDH; E3 component of pyruvate dehydrogenase complex, 2-oxo-glutarate complex; E3; EC 1.8.1; EC 1.8.1.4; GCSL; Glycine cleavage system L protein; glycine cleavage system protein L; LAD; Lipoamide Dehydrogenase; lipoamide reductase; lipoyl dehydrogenase; PHE3