Anti-Pyruvate Dehydrogenase Immunocapture antibody [15D3G9C11] (ab109866)

Overview

  • Product nameAnti-Pyruvate Dehydrogenase Immunocapture antibody [15D3G9C11]
  • Description
    Mouse monoclonal [15D3G9C11] to Pyruvate Dehydrogenase Immunocapture
  • Specificity100 µg ab109866 can capture at least 5 µg Pyruvate Dehydrogenase complex from 1 mg solubilized Bovine heart mitochondria.
  • Tested applicationsSuitable for: ICC/IF, Flow Cyt, IPmore details
    Unsuitable for: IHC-P or WB
  • Species reactivity
    Reacts with: Cow, Human
    Predicted to work with: Pig
  • Immunogen

    Pig heart Pyruvate Dehydrogenase

  • Positive control
    • Bovine heart mitochondria This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.
  • General notes

    Product was previously marketed under the MitoSciences sub-brand.

Properties

Applications

Our Abpromise guarantee covers the use of ab109866 in the following tested applications.

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

Application Abreviews Notes
ICC/IF Use a concentration of 10 µg/ml.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IP Use at 100-1 µg/mg of lysate.
  • Application notesIs unsuitable for IHC-P or WB.
  • Target

    • RelevanceThe pyruvate dehydrogenase complex (PDH) is at the center of aerobic carbohydrate metabolism. It is localized in the matrix space of mitochondria where it catalyzes the irreversible oxidative decarboxylation of pyruvate entering the organelle to produce acetyl-CoA, NADH and CO2. PDH is a complex of three different enzymes that channel substrate and product. These are pyruvate dehydrogenase (E1), dihydrolipoamide transacetylase (E2) and dihydrolipoamide dehydrogenase (E3). PDH deficiencies, diabetes, starvation, sepsis, and possibly Alzheimer's disease have been linked to altered PDH functioning.
    • Cellular localizationMitochondrial

    Anti-Pyruvate Dehydrogenase Immunocapture antibody [15D3G9C11] images

    • ICC/IF image of ab109866 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 ab109866 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) 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.

    • Immunoprecipitation of Pyruvate Dehydrogenase using ab109866 at 100 µg/mg lysate.
    • Overlay histogram showing HepG2 cells stained with ab109866 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab109866, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    References for Anti-Pyruvate Dehydrogenase Immunocapture antibody [15D3G9C11] (ab109866)

    This product has been referenced in:
    • Murray J  et al. Focused proteomics: towards a high throughput monoclonal antibody-based resolution of proteins for diagnosis of mitochondrial diseases. Biochim Biophys Acta 1659:206-11 (2004). Read more (PubMed: 15576053) »

    See 1 Publication for this product

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    Thank you for your call yesterday and for your patience while I have been in touch with the lab regarding your enquiry.

    The immunocapture antibody ab109866 is actually our PDH E2/E3bp antibody ab110333 conjugated to beads for IP. So, ab109866...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"