ZNF420 Antibody, FITC conjugated

Introduction to ZNF420 Antibody, FITC Conjugated

ZNF420 is a transcription factor containing 19 C2H2-type zinc fingers and a KRAB domain, implicated in transcriptional regulation and p53-mediated apoptosis . The FITC-conjugated ZNF420 antibody enables direct visualization of this protein in cellular and tissue samples, leveraging fluorescein’s fluorescent properties for techniques like immunofluorescence (IF) and immunohistochemistry (IHC).

FITC Conjugation and Performance

• Labeling Impact: Higher FITC-to-antibody ratios reduce binding affinity due to steric hindrance but increase sensitivity in low-abundance targets .

• Optimal Conjugation: Achieved at pH 9.5, 25 mg/mL antibody concentration, and 30–60 minutes at room temperature, yielding an F/P (fluorescein-to-protein) ratio of 2–4 .

• Non-Specific Staining: Over-conjugated antibodies (>6 FITC molecules per IgG) show increased background noise .

Functional Insights

• Biological Role: ZNF420 acts as a negative regulator of p53 under non-stress conditions. Phosphorylation by ATM during DNA damage dissociates ZNF420 from p53, enabling apoptosis .

• Chromatin Organization: ZNF420 interacts with chromatin-modifying complexes, influencing gene expression in development and cancer .

Applications and Validation

• Immunofluorescence: Validated in cultured cells and paraffin-embedded mouse tissues, showing nuclear localization of ZNF420 .

• Cross-Reactivity: Confirmed reactivity with human, rat, and cow homologs via sequence alignment .

• Controls Required: Include unstained samples and isotype-matched antibodies to mitigate Proclin 300-induced autofluorescence .

Comparative Analysis of ZNF420 Antibody Conjugates

Critical Considerations

• Preservative Hazards: Proclin 300 in the storage buffer is toxic; handle with PPE .

• Batch Variability: Assess F/P ratios via absorbance (A495/A280) before use .

• Alternative Conjugates: For multiplex imaging, consider AbBy Fluor® 488 or 555 to avoid spectral overlap .