Target Biology: Pyruvate Dehydrogenase E2 Subunit

The pyruvate dehydrogenase complex (PDC) is a large multi-enzyme assembly located in the mitochondrial matrix that catalyzes the irreversible oxidative decarboxylation of pyruvate to acetyl-CoA, thereby linking cytoplasmic glycolysis to the mitochondrial tricarboxylic acid (TCA) cycle. The E2 subunit, encoded by the DLAT gene (dihydrolipoamide S-acetyltransferase), serves as the structural and catalytic core of the complex. It contains lipoyl domains that facilitate the transfer of reaction intermediates among the three catalytic components: E1 (pyruvate decarboxylase), E2 (dihydrolipoamide acetyltransferase), and E3 (dihydrolipoamide dehydrogenase).

Dysregulation of PDC activity and mutations in DLAT have been associated with metabolic disorders including pyruvate dehydrogenase deficiency, and PDC components are frequently studied in the context of cancer metabolism, neurodegeneration, and autoimmune disease such as primary biliary cholangitis (PBC), where anti-mitochondrial antibodies targeting the E2 subunit are a hallmark diagnostic feature.

Antibody Format and Formulation

Ab350126 is a mouse-derived monoclonal antibody raised against the Pyruvate Dehydrogenase E2 subunit. Monoclonal antibodies offer a single, defined specificity, ensuring high lot-to-lot reproducibility that is critical for quantitative and comparative experiments. The antibody is referenced to the absea clone KC-956, providing a traceable source lineage for regulatory and publication purposes.

The product is supplied in phosphate-buffered saline (PBS) only, without additional carrier proteins or preservatives unless otherwise specified on the certificate of analysis. Researchers should confirm final formulation details—including concentration, stabilizers, and storage conditions—with the product datasheet or certificate of analysis prior to use. PBS-formulated antibodies are compatible with direct conjugation workflows and downstream applications requiring protein-free conditions.

Research Applications

Mouse monoclonal antibodies targeting the PDH E2 subunit are employed across a range of laboratory techniques. Common applications include:

• Western Blotting (WB): Detection of DLAT protein expression levels in cell and tissue lysates, with an expected band at approximately 70 kDa.
• Immunofluorescence (IF) / Immunocytochemistry (ICC): Visualization of the mitochondrial localization pattern of PDH E2, useful for studying mitochondrial network dynamics.
• Immunohistochemistry (IHC): Assessment of PDH E2 expression in formalin-fixed, paraffin-embedded (FFPE) tissue sections for pathological studies.
• Immunoprecipitation (IP): Pull-down of the PDC or DLAT-interacting protein networks for proteomic analysis.
• ELISA: Quantitative detection of PDH E2 in cell extracts or as a capture/detection antibody in sandwich assay formats.

Optimal working dilutions for each application should be determined empirically. Researchers are advised to consult the product datasheet for validated application data specific to Ab350126.

Storage, Handling, and Quality Considerations

Proper storage and handling of monoclonal antibodies are essential to maintain activity and specificity. General best practices for PBS-formulated antibodies include:

• Store at 2–8 °C for short-term use (up to one month) or at −20 °C for long-term storage. Avoid repeated freeze-thaw cycles, which can cause aggregation and loss of activity.
• Centrifuge briefly before opening to collect the solution at the bottom of the vial.
• Aliquot into single-use volumes for long-term storage to minimize freeze-thaw events.
• Keep away from light to prevent potential photodegradation of any conjugated labels if applicable.

Note: Specific storage recommendations, shelf life, and concentration details for Ab350126 are provided on the product datasheet and certificate of analysis. Always refer to these documents for authoritative guidance.