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Signal Transduction Metabolism Amino Acids
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

  • Datasheet
Reviews (1) Submit a question References (5)

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Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • OI-RD Scanning - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
  • Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6635] to Lipoamide Dehydrogenase
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-Lipoamide Dehydrogenase antibody [EPR6635]
    See all Lipoamide Dehydrogenase primary antibodies
  • Description

    Rabbit monoclonal [EPR6635] to Lipoamide Dehydrogenase
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to residues in Human Lipoamide Dehydrogenase (UniProt ID: P09622).

  • Positive control

    • WB: Jurkat, HeLa, 293T, MCF7, and Caco-2 cell lysates. IHC-P: Human kidney and Human colon tissues
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 1.52 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.2
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR6635
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Amino acid metabolism

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab206723)
    • Anti-Lipoamide Dehydrogenase antibody [EPR6635] - BSA and Azide free (ab240067)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human DLD (Lipoamide Dehydrogenase) knockout HeLa cell line (ab265535)
  • KO cell lysates

    • Human DLD (Lipoamide Dehydrogenase) knockout HeLa cell lysate (ab257922)
  • Positive Controls

    • MCF7 whole cell lysate (ab29537)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • Caco-2 whole cell lysate (ab3950)
    • Jurkat whole cell lysate (ab7899)
  • Recombinant Protein

    • Recombinant Human Lipoamide Dehydrogenase protein (ab93679)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133551 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Human
WB
Human
All applications
Mouse
Rat
Application Abreviews Notes
WB (1)
1/10000 - 1/50000. Detects a band of approximately 56 kDa (predicted molecular weight: 54 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/10000 - 1/50000. Detects a band of approximately 56 kDa (predicted molecular weight: 54 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Function

      Lipoamide dehydrogenase is a component of the glycine cleavage system as well as of the alpha-ketoacid dehydrogenase complexes. Involved in the hyperactivation of spermatazoa during capacitation and in the spermatazoal acrosome reaction.
    • Involvement in disease

      Note=Defects in DLD are involved in the development of congenital infantile lactic acidosis.
      Defects in DLD are a cause of maple syrup urine disease (MSUD) [MIM:248600]. MSUD is characterized by mental and physical retardation, feeding problems and a maple syrup odor to the urine. The keto acids of the branched-chain amino acids are present in the urine, resulting from a block in oxidative decarboxylation.
    • Sequence similarities

      Belongs to the class-I pyridine nucleotide-disulfide oxidoreductase family.
    • Post-translational
      modifications

      Tyrosine phosphorylated.
    • Cellular localization

      Mitochondrion matrix.
    • Target information above from: UniProt accession P09622 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 1738 Human
      • Entrez Gene: 13382 Mouse
      • Entrez Gene: 298942 Rat
      • Omim: 238331 Human
      • SwissProt: P09622 Human
      • SwissProt: O08749 Mouse
      • SwissProt: Q6P6R2 Rat
      • Unigene: 131711 Human
      • Unigene: 3131 Mouse
      • Unigene: 86962 Rat
      see all
    • Alternative names

      • Dehydrogenase complex, E3 component antibody
      • Diaphorase antibody
      • Dihydrolipoamide dehydrogenase antibody
      • Dihydrolipoyl dehydrogenase antibody
      • Dihydrolipoyl dehydrogenase mitochondrial antibody
      • dld antibody
      • DLDD antibody
      • DLDH antibody
      • DLDH_HUMAN antibody
      • E3 antibody
      • E3 branched chain aplha-keto acid antibody
      • E3 component of pyruvate dehydrogenase antibody
      • E3 component of pyruvate dehydrogenase complex 2 oxo glutarate complex branched chain keto acid dehydrogenase complex antibody
      • GCSL antibody
      • Glycine cleavage system L protein antibody
      • Glycine cleavage system protein L antibody
      • LAD antibody
      • lipoamide dehydrogenase antibody
      • Lipoamide reductase antibody
      • Lipoyl dehydrogenase antibody
      • mitochondrial antibody
      • OTTHUMP00000206744 antibody
      • OTTHUMP00000206746 antibody
      • OTTHUMP00000206748 antibody
      • OTTHUMP00000206749 antibody
      • PHE 3 antibody
      • PHE3 antibody
      • Pyruvate dehydrogenase component E3 antibody
      see all

    Images

    • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      All lanes : Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : DLD knockout HeLa cell lysate
      Lane 3 : Jurkat cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 54 kDa
      Observed band size: 56 kDa
      why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab133551 observed at 56 kDa. Red - loading control ab8245 observed at 36 kDa.

      ab133551 Anti-Lipoamide Dehydrogenase antibody [EPR6635] was shown to specifically react with Lipoamide Dehydrogenase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265535 (knockout cell lysate ab257922) was used. Wild-type and Lipoamide Dehydrogenase knockout samples were subjected to SDS-PAGE. ab133551 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

      Immunohistochemical analysis of paraffin embedded Human colon tissue labelling Lipoamide Dehydrogenase with ab133551 antibody at a dilution of 1/100.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      All lanes : Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551) at 1/10000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : 293T cell lysate
      Lane 4 : MCF7 cell lysate
      Lane 5 : Caco-2 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 54 kDa
      Observed band size: 56 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

      Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling Lipoamide Dehydrogenase with ab133551 antibody at a dilution of 1/100.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • OI-RD Scanning - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      OI-RD Scanning - Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)
      Anti-Lipoamide Dehydrogenase antibody [EPR6635] (ab133551)

    Protocols

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
  • References (5)

    Publishing research using ab133551? Please let us know so that we can cite the reference in this datasheet.

    ab133551 has been referenced in 5 publications.

    • Dobolyi A  et al. Exclusive neuronal detection of KGDHC-specific subunits in the adult human brain cortex despite pancellular protein lysine succinylation. Brain Struct Funct 225:639-667 (2020). PubMed: 31982949
    • Tang M  et al. Hippocampal proteomic changes of susceptibility and resilience to depression or anxiety in a rat model of chronic mild stress. Transl Psychiatry 9:260 (2019). PubMed: 31624233
    • Bugarski M  et al. Multiphoton imaging reveals axial differences in metabolic autofluorescence signals along the kidney proximal tubule. Am J Physiol Renal Physiol 315:F1613-F1625 (2018). PubMed: 30132348
    • Ferriero R  et al. Phenylbutyrate increases pyruvate dehydrogenase complex activity in cells harboring a variety of defects. Ann Clin Transl Neurol 1:462-70 (2014). PubMed: 25356417
    • Roselló-Lletí E  et al. Heart mitochondrial proteome study elucidates changes in cardiac energy metabolism and antioxidant PRDX3 in human dilated cardiomyopathy. PLoS One 9:e112971 (2014). WB ; Human . PubMed: 25397948

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Western blot abreview for Anti-Lipoamide Dehydrogenase antibody [EPR6635]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - other (Striatal cell line, pheochromocytoma cell line)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris gradient gel. Laemmli buffer with 10% β-Mercaptoethanol. Samples were denatured at 55C for 10 min. Immobilon-FL transfer membranes. Overnight transfer at +4C.)
    Loading amount
    20 µg
    Specification
    Striatal cell line, pheochromocytoma cell line
    Blocking step
    SEA Block Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 04 2018

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