Anti-COX5B Rabbit Monoclonal Antibody for WB - P10606

Core Product Specifications and Parameters

Biological Background: COX5B Function and Localization

• COX5B (cytochrome c oxidase subunit 5B) is a nuclear-encoded subunit of mitochondrial complex IV (cytochrome c oxidase), the terminal enzyme of the electron transport chain.
• It is a component of the multi-subunit cytochrome c oxidase complex that catalyzes the reduction of molecular oxygen to water, coupling this reaction to proton pumping across the inner mitochondrial membrane.
• The COX5B protein is synthesized with a mitochondrial transit peptide, which is cleaved after import into the mitochondrion, yielding a mature subunit that integrates into the inner membrane.
• Subcellular localization is restricted to the mitochondrial inner membrane, where it contributes to the structural and functional integrity of complex IV.
• Post-translational modifications include acetylation, and the mature protein contains a zinc-binding site, as indicated by the presence of a zinc ion in the crystal structure.
• The theoretical molecular weight of the mature human COX5B is approximately 13.7 kDa, consistent with the observed 13 kDa band on SDS-PAGE.
• As a core subunit of the respiratory chain, COX5B is essential for efficient oxidative phosphorylation and ATP production, and its expression is often used as a marker for mitochondrial mass and activity.

Experimental Guidance and Technical Tips

• The immunogen spans amino acids 29–129 of human COX5B, which corresponds to the mature processed protein lacking the mitochondrial targeting peptide, likely generating antibodies that recognize the endogenous processed form.
• For Western blotting, a band around 13 kDa is expected in lysates from human, mouse, and rat mitochondria-rich tissues (e.g., heart, skeletal muscle, liver). Use a mitochondrial fraction enriched sample for optimal detection.
• For immunohistochemistry (IHC-P) and immunofluorescence (IF/ICC), staining should show a punctate cytoplasmic pattern consistent with mitochondrial localization. Counterstaining with a mitochondrial marker is recommended for validation.
• In immunoprecipitation (IP) experiments, the antibody may co-precipitate other complex IV subunits; consider using mild detergents to preserve native complex integrity.
• ELISA reactivity may be used for quantitative detection of COX5B in cell or tissue lysates, but performance should be validated with known standards.
• Cross-reactivity with mouse and rat has been confirmed, but always verify species-specific reactivity in your own experimental system due to potential sequence variations.

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