Product nameAnti-pan PDK antibody [3H5BE12]
DescriptionMouse monoclonal [3H5BE12] to pan PDK
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Human
Recombinant full length protein
This antibody clone is manufactured by Abcam.
PDH Kinases (PDKs) deactivate PDH by reversible phosphorylation. In humans and rodents, 4 PDK's are present in an organ-dependent manner. In all cases, PDK's are physically part of the PDH complex (binding via lipoyl domain 2 of E2), and they phosphorylate the E1 subunit at 3 specific serine residues. PDH phosphatases remove the phosphorylation and restore activity of PDH.
Product was previously marketed under the MitoSciences sub-brand.
Storage instructionsShipped at 4°C. Store at +4°C.
Storage bufferPreservative: 0.02% Sodium azide
Constituent: HEPES buffered saline
Concentration information loading...
Purity>95% by SDS-PAGE
Purification notesNear homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
Primary antibody notesPDH Kinases (PDKs) deactivate PDH by reversible phosphorylation. In humans and rodents, 4 PDK's are present in an organ-dependent manner. In all cases, PDK's are physically part of the PDH complex (binding via lipoyl domain 2 of E2), and they phosphorylate the E1 subunit at 3 specific serine residues. PDH phosphatases remove the phosphorylation and restore activity of PDH.
Light chain typekappa
Our Abpromise guarantee covers the use of ab115321 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 49 kDa.|
All lanes : Anti-pan PDK antibody [3H5BE12] (ab115321) at 1 µg/ml
Lane 1 : human heart mitochondria at 20 µg
Lane 2 : human liver mitochondria at 10 µg
Lane 3 : rat liver mitochondria at 10 µg
Lane 4 : mouse liver mitochondria at 10 µg
Lane 5 : PDK1 recombinant protein ab110359
at 0.1 µg
Lane 6 : PDK2 recombinant protein ab110354 at 0.1 µg
Lane 7 : PDK3 recombinant protein ab110355 at 0.1 µg
Lane 8 : PDK4 recombinant protein ab110356 at 0.1 µg
Lane 9 : PDK1 recombinant protein ab86986 at 0.1 µg
All lanes : GAM IR 800 1:5000
Performed under reducing conditions.
Predicted band size: 49 kDa
ab115321 identifies all four isoforms of PDKs (PDK1, PDK2, PDK3, and PDK4) with differential affinity. PDK3 has the highest affinity, then PDK2 and 4, it has the lowest affinity to PDK1.
IHC image of ab115321 staining in human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab115321, 5µg/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.
Western blot analysis of human ovarian cancer cell lines whole cell lysate (30µg/lane) labelling pan PDK with ab115321 at 1/1000. A HRP-conjugated goat anti-mouse IgG polyclonal (1/10000) was used as the secondary antibody.