SPEF2 Antibody, FITC conjugated

Definition and Purpose

The SPEF2 antibody, FITC conjugated is a fluorescently labeled immunoglobulin designed to detect the Sperm Flagellar 2 (SPEF2) protein. SPEF2, also known as Kpl2, is critical for ciliary function and spermatogenesis . The FITC (Fluorescein Isothiocyanate) conjugation enables visualization of SPEF2 via fluorescence microscopy, particularly in immunofluorescence (IF) assays .

Antibody Characteristics

Data compiled from .

Conjugation Process

FITC binds covalently to primary amines (e.g., lysine residues) on the antibody. Optimal conjugation ratios balance fluorescence brightness and antibody solubility:

• FITC-to-protein ratio: 3–6 FITC molecules per antibody .

• Reaction conditions:

  • pH: 9.0–9.5 (optimal for isothiocyanate reactivity) .

  • Temperature: Room temperature (20–25°C) .

  • Time: 1–2 hours for completion .

Critical Considerations

• Sodium azide removal: Required before conjugation to avoid FITC inactivation .

• Light sensitivity: FITC degrades under prolonged light exposure; store at -20°C in dark conditions .

Role of SPEF2 in Biological Systems

SPEF2 is essential for:

• Ciliary function: Mutations in Spef2 cause immotile cilia and primary ciliary dyskinesia (PCD) in mice .

• Spermatogenesis: Required for sperm flagellar biogenesis; defects lead to infertility .

• Pancreatic signaling: Interacts with AMPKα/Sirt1 pathways to modulate acinar cell responses in pancreatitis .

Experimental Uses of SPEF2-FITC Antibody

Recommended Dilution

• Immunofluorescence: 1:500 in PBS + 10% FBS .

Key Challenges

• Cross-reactivity: Limited to human samples; species-specific validation required .

• Background noise: Higher FITC conjugation ratios may increase nonspecific binding .

Comparison with Related Antibodies

Data derived from .