CUP1-1 Antibody, FITC conjugated

FITC Conjugation Details

The antibody is conjugated to fluorescein isothiocyanate (FITC), a fluorescent dye with excitation/emission maxima at 499/515 nm . The conjugation process involves:

1. Dialysis to remove impurities and adjust pH to 9.2–9.5 .

2. Reaction with FITC (5–10 mg/ml in anhydrous DMSO) for 2–4 hours at room temperature .

3. Purification via gel filtration or DEAE Sephadex chromatography to remove unbound FITC .

Critical considerations:

• Higher FITC labeling indices correlate with reduced antibody binding affinity but increased sensitivity in immunohistochemistry .

• Optimal labeling conditions include pH 9.5, 25 mg/ml protein concentration, and 30–60 minutes reaction time .

Applications

Research Findings

• CUP1-1 Function: The antibody targets a protein critical for copper detoxification and storage in yeast, regulating oxidative stress responses .

• Cancer Research: CUP1-1 homologs in humans (e.g., metallothioneins) are implicated in tumor progression and chemotherapy resistance .

• Optimization Studies: FITC-conjugated antibodies with lower labeling indices (F/P ratio <3) exhibit higher specificity in TCR studies .