PRAMEF6 Antibody, HRP conjugated

Introduction to PRAMEF6 Antibody, HRP Conjugated

The PRAMEF6 Antibody, HRP conjugated is a specialized reagent used in molecular biology and immunology research. PRAMEF6 (PRAME Family Member 6) is a substrate-recognition component of the Cul2-RING E3 ubiquitin-protein ligase complex, mediating ubiquitination of truncated selenoproteins like MSRB1/SEPX1 . The HRP (Horseradish Peroxidase) conjugation enables detection via chromogenic or chemiluminescent substrates, commonly used in ELISA, western blotting, and immunohistochemistry (IHC). This article synthesizes data from diverse sources to provide a comprehensive overview of its structure, conjugation methods, applications, and research findings.

Structure and Function of PRAMEF6

• Biological Role: PRAMEF6 facilitates ubiquitination of selenoproteins, targeting them for degradation when translation termination is impaired .

• Antibody Specificity: The antibody targets the C-terminal region (AA 395–423) of human PRAMEF6, ensuring specificity for this isoform .

• Conjugation: HRP is covalently linked to the antibody’s lysine residues, retaining enzymatic activity for detection .

Conjugation Methods

HRP conjugation is achieved through three primary methods:

Traditional Conjugation

• Process: Cross-linking via NHS-ester or maleimide chemistry .

• Advantage: High flexibility in antibody selection.

• Limitation: Requires purification and optimization.

Lightning-Link® HRP Kit

• Process: Pre-activated HRP reacts with antibody lysines in 2–3 hours .

• Advantage: No purification needed; includes LifeXtend™ stabilizer for long-term storage.

oYo-Link® HRP

• Process: Site-specific labeling of antibody heavy chains via photocrosslinking .

• Advantage: Rapid conjugation (30 seconds hands-on, 2 hours total); uniform labeling (2 HRP/antibody).

ELISA

• Dilution: 0.7–200 ng/mL (dependent on target concentration) .

• Detection: TMB or ABTS substrates generate colorimetric signals .

Western Blotting

• Dilution: 1:1,000–1:5,000 .

• Detection: Enhanced chemiluminescence (ECL) with HRP substrate .

Immunohistochemistry (IHC)

• Dilution: 1:10–1:50 .

• Detection: DAB (brown precipitate) or TrueBlue (purple) .

Flow Cytometry

• Dilution: 1:10–1:50 .

• Use: Intracellular staining of PRAMEF6 in fixed/permeabilized cells.

Research Findings

• Cancer Research: PRAMEF6 expression is linked to tumor progression in melanoma and ovarian cancer .

• Neurodegeneration: Its role in selenoprotein degradation suggests potential in studying neurodegenerative diseases .

• Immunoassay Optimization: HRP-conjugated antibodies reduce cross-reactivity compared to secondary antibodies .