BBC3 Antibody, HRP conjugated

Compound Overview

BBC3 Antibody, HRP conjugated is a high-quality polyclonal antibody raised in rabbits against a synthetic peptide derived from human PUMA (Q96PG8) . The HRP conjugation facilitates enzymatic signal amplification in immunoassays, allowing quantitative or qualitative detection of BBC3, a pro-apoptotic protein involved in p53-mediated apoptosis .

Applications and Performance

The conjugate is optimized for ELISA, where HRP catalyzes colorimetric or chemiluminescent reactions upon substrate addition (e.g., TMB or ABTS) . Key features include:

• High sensitivity: HRP amplifies signal output, enabling detection of low-abundance BBC3.

• Specificity: Targets the mitochondrial BBC3 protein, critical for apoptosis regulation .

• Stability: Glycerol in the formulation prevents aggregation during storage .

Conjugation Methodology

While the exact protocol for this specific conjugate is undisclosed, HRP-antibody conjugation typically involves:

1. Antibody Modification: Introducing thiol groups (-SH) to IgG antibodies using heterobifunctional crosslinkers (e.g., maleimide-activated HRP) .

2. HRP Activation: Periodate oxidation of HRP’s carbohydrate moieties to generate aldehyde groups for amine coupling .

3. Purification: Removal of unreacted HRP/antibody via size-exclusion chromatography or scavenger resins to achieve >90% purity .

Critical Parameters for Conjugation

• Antibody Purity: >90% pure IgG by SEC-HPLC or gel electrophoresis .

• HRP Quality: >99% purity and >200 units/mg enzymatic activity .

• Stoichiometry: Target 2–4 HRP molecules per antibody to avoid polymerization .

Research Insights

Recent advancements in HRP-antibody conjugation highlight methods to improve assay sensitivity:

• Lyophilization: Freeze-drying activated HRP increases antibody-HRP binding efficiency, enhancing ELISA sensitivity by ~200-fold compared to classical methods .

• Recombinant Conjugates: Genetic fusion of HRP with antibody fragments (e.g., Fab) in Pichia pastoris yields homogeneous conjugates with retained antigen-binding and enzymatic activity .

Comparative Analysis

Future Directions

• Enhanced Stability: Reducing glycerol dependency through lyophilization .

• Multiplexing: Combining BBC3-HRP with other reporter enzymes (e.g., alkaline phosphatase) for dual-target assays.