NDUFB8 Antibody, HRP conjugated

Target Protein: NDUFB8

NDUFB8 (NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 8) functions as an accessory subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase, also known as Complex I. This protein is believed not to be directly involved in catalysis but serves important structural roles within the complex. Complex I plays a crucial role in transferring electrons from NADH to the respiratory chain, with ubiquinone functioning as the immediate electron acceptor for the enzyme . NDUFB8 is commonly used as a marker for mitochondrial function in various research contexts and clinical investigations, particularly in studies examining mitochondrial disorders.

Antibody Structure and Conjugation

NDUFB8 antibodies conjugated with HRP are available in various forms, including both polyclonal and monoclonal variants. The horseradish peroxidase (HRP) enzyme is covalently linked to the antibody molecule, enabling direct enzymatic detection without requiring secondary antibody steps. This conjugation provides significant advantages in terms of assay simplicity, reduced background, and enhanced signal detection capabilities. Commercial preparations typically contain stabilizers and preservatives to maintain antibody activity during storage and use .

Available Variants and Sources

Multiple vendors offer NDUFB8 antibody, HRP conjugated products with varying specifications. These include recombinant and conventional antibody options, with differences in host species, clonality, and epitope targeting. The most common variants are derived from rabbit hosts, targeting specific amino acid sequences of the human NDUFB8 protein .

Target Protein Characteristics

NDUFB8 has specific molecular identifiers in protein databases that help researchers correctly identify and work with this target:

Table 2: NDUFB8 protein characteristics and identifiers

Validated Applications

NDUFB8 antibody, HRP conjugated products have been validated for multiple research applications with varying dilution recommendations:

Table 3: Recommended dilutions for various applications

Multiplex Immunofluorescence Applications

NDUFB8 antibody has been specifically optimized for use in multiplex immunofluorescence assays, where multiple markers are detected simultaneously. Research has shown that NDUFB8 antibody-HRP conjugates perform effectively in such systems when appropriate signal amplification strategies are employed. Due to the relatively low abundance of NDUFB8 protein in many samples, enhanced detection systems are often necessary for optimal results .

Signal Amplification Strategies

Research has demonstrated that standard OmniMap HRP amplification may be inadequate for NDUFB8 detection due to the low abundance of this target antigen. To address this limitation, enhanced signal amplification protocols have been developed. One effective approach involves using a secondary antibody conjugated to hapten-based anti-mouse HQ polymer followed by an HRP-conjugated anti-HQ tertiary antibody. This multi-step amplification significantly improves NDUFB8 fluorescence signal intensity in multiplex assays .

Denaturation Considerations

Studies have shown that NDUFB8 antibody-HRP conjugates undergo robust heat-mediated denaturation under standard conditions (CC2 95°C for 8 minutes). This characteristic makes NDUFB8 staining particularly suitable as an early cycle in multiplex staining protocols. When designing multiplex assays that include NDUFB8 antibody, HRP conjugated, researchers have found that positioning the NDUFB8 cycle first in the staining sequence is advantageous. This placement helps minimize potential degradation of the target antigen during subsequent heat-mediated denaturation steps required for other markers .

Multiplex Protocol Example

The following table presents an optimized multiplex protocol incorporating NDUFB8 antibody, HRP conjugated:

Table 4: Optimized multiplex staining protocol incorporating NDUFB8 antibody

Reproducibility Assessment

Quantitative single-cell analysis has confirmed the effective denaturation of NDUFB8 antibody-HRP complexes in automated multiplex assays. Research comparing staining results across multiple batches using serial sections from tissue samples has demonstrated high reproducibility of signal intensity. This reproducibility represents a key advantage of automated approaches for NDUFB8 detection, particularly in clinical and diagnostic applications where consistency is critical .

Cross-Reactivity Profiles

Different commercial preparations of NDUFB8 antibody, HRP conjugated exhibit varying cross-reactivity profiles. While some products are highly specific for human NDUFB8, others demonstrate broader cross-reactivity with mouse and rat orthologs. Researchers should carefully select appropriate antibody products based on their specific experimental models and cross-reactivity requirements .

Quality Control Considerations

When working with NDUFB8 antibody, HRP conjugated, researchers should implement appropriate controls to ensure specific staining and accurate interpretation of results. These include negative controls (isotype-matched control antibodies) to assess non-specific binding and positive controls (tissues or cell lines known to express NDUFB8) to confirm detection sensitivity. For multiplex applications, adjacent sections labeled with IgG control antibodies should be used to assess background signal .