Anti-HIBADH Monoclonal Antibody for WB, IHC-P, IF/ICC, ELISA - P31937

Core Product Specifications and Parameters

Biological Background: HIBADH Function and Localization

• HIBADH (3-hydroxyisobutyrate dehydrogenase) is a mitochondrial enzyme belonging to the oxidoreductase family, utilizing NAD as a cofactor.
• It plays a key role in the catabolism of branched-chain amino acids (valine, leucine, isoleucine), specifically in the conversion of 3-hydroxyisobutyrate to methylmalonate semialdehyde.
• The protein is localized to the mitochondrion, as indicated by its mitochondrial transit peptide, which is cleaved to yield the mature enzyme.
• Post-translational modifications include acetylation, which may regulate its enzymatic activity or stability.
• HIBADH expression has been detected in skin fibroblasts; its expression in other tissues may be inferred from its metabolic role.
• Structural studies have provided a 3D-structure for HIBADH, aiding in understanding its catalytic mechanism and substrate recognition.
• The protein has been identified through direct protein sequencing and proteomics approaches, confirming its presence in human mitochondria.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 37-336 of human HIBADH (NP_689953.1), which excludes the predicted mitochondrial transit peptide (approximately the first 36 amino acids); the antibody is therefore likely to recognize the mature mitochondrial form.
• For Western blot (WB), a band at approximately 35 kDa is expected (consistent with UniProt molecular weight of 35.3 kDa). Confirm specificity by using appropriate positive and negative controls (e.g., HIBADH-overexpressing cells or tissues with known expression).
• In immunohistochemistry (IHC-P) and immunofluorescence (IF/ICC), mitochondrial staining patterns should be observed. Co-localization with mitochondrial markers can help validate specificity.
• The antibody cross-reacts with human, mouse, and rat HIBADH; when using rodent samples, verify sequence homology in the immunogen region to ensure comparable reactivity.
• For ELISA applications, consider using the recombinant immunogen as a positive control and titrate the antibody concentration for optimal signal-to-noise ratio.
• To minimize non-specific binding, suitable blocking buffers (e.g., 5% non-fat milk or BSA) and appropriate dilution optimization are recommended.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio supports diagnostic manufacturers and researchers by providing validated antibody pairs, optimized monoclonal/polyclonal antibodies, bulk ancillary reagents, and rare target raw-material sourcing. For metabolic and mitochondrial disease research, CamelBio offers HIBADH-specific reagents and customizable raw-material solutions to accelerate assay development. Our one-stop supply chain ensures seamless transition from concept to clinical application.