EnkiLife R-PE labeling kit contains multiple components for labeling antibody proteins, such as the activated lyophilized R-PE fluorescent dye, labeling buffer, and other chemical reagents, enabling easy and reliable antibody labeling for use as fluorescent probes.

Features

lActivated dye with extended arm: A specially designed long flexible arm minimizes spatial hindrance between antibodies and the fluorescent dye. This protects antibody activity and enhances labeling efficiency.

lSite-specific conjugation: Optimized site-specific conjugation technology ensures uniform antibody labeling and improves efficiency.

lRapid process: The simple procedure allows for efficient labeling completion in as fast as 3 - 4 hours.

lHigh dye brightness: The selected high-performance fluorescent dye boasts high quantum yield and brightness, ensuring high detection sensitivity.

lDye stability: The chosen high-performance fluorescent dye is stable, resistant to degradation, and suitable for various subsequent applications, such as fixed sample detection.

lUser-friendly operation: Each dye reagent is pre-optimized for corresponding antibody amounts. No complex calculations or ultrafiltration steps are required if the antibody sample meets the labeling conditions. The solid dye form ensures batch stability. Simply mix the reagents according to the steps to achieve good results.

lFlexibility: In the large-scale kit, the dye is pre-portioned for consistent batch performance, facilitating multiple labeling uses.

lComprehensive reagents: The kit includes a labeling preservation solution, covering the entire process from labeling to preservation. No additional reagents are needed, making the process worry-free and convenient.

Dye Introduction

R-Phycoerythrin (R-PE) is a strong fluorescent phycobiliprotein isolated from red algae. With an extremely high quantum yield, it emits bright orange - red fluorescence. As an excellent fluorescent labeling dye, it is widely used in biological detection. Note that phycobiliproteins mainly exist in two forms: R-PE and B-PE, which have some differences in optical properties. This kit uses R-PE sourced from red algae. Its main absorption peak is at 565 nm, with minor peaks at 496 nm and 545 nm. The prominence of the minor peaks varies significantly across different species. PE consists of three subunits: α (20,000 Da), β (20,000 Da), and γ (30,000 Da). The molecular weight of intact PE is approximately 240,000 Da, with a quantum efficiency exceeding 90%. Its orange fluorescence is easily visible to the naked eye in most solutions.

Properties of R-PE Dye

Synonyms: PE, Phycoerythrin

Form: Red solid

Molecular Weight: ~ 240 kDa

Excitation Light Sources: 488 nm, 532 nm, 561 nm

Excitation Wavelengths: 496 nm, 566 nm

Emission Wavelength: 576 nm

Molar Extinction Coefficient (L/(mol・cm)): 1960000

Purity: A566/A280 ≥ 5.3

Solubility: Highly soluble in aqueous solutions

Activating Group: Maleimide (MAL)

Activation Effect: MAL/R-PE = 1.5 - 4

R-PE Dye Spectrum

Kit Specifications

Examples of R-PE Dye - Labeled Antibody/Protein Applications

nR-PE - labeled anti - Human CD25 for immunophenotyping (human peripheral blood samples). Detection marker combinations like CD4 - FITC + CD8 - APC + CD25 - R-PE (IL - 2 receptor, a marker for regulatory T cells) enable precise identification of low - expressed CD25 targets, preventing missed detection of weak signals.

nR-PE - labeled anti - CD68 in a red - green combination (anti - PD - L1 Fluor 488 + CD68 - R-PE + DAPI) reduces channel crosstalk and effectively calculates the spatial co - localization rate of CD68 - positive macrophages and PD - L1 - positive cells in the tumor stroma.

nCoating autoantigens (e.g., dsDNA) → adding patient serum → using R-PE - labeled anti - human IgG secondary antibodies in autoimmune antibody test kit development. Fluorescent enzyme - labeled detection improves assay sensitivity.

nEGFR - R-PE antibodies → binding to cell surfaces at 4°C → activation of internalization at 37°C → flow cytometry detects signal attenuation, enabling tracking of antibody - mediated endocytosis.

nStreptavidin - R-PE + Biotinylated antibodies enhance detection rates of weakly expressed targets (e.g., CTLA - 4) by 30 times.

nApplications in liquid - phase arrays enable simultaneous detection of 12 cytokines in a single well (different microsphere encoding + R-PE - labeled detection antibodies with unified signals) for multi - analyte detection.

Labeling Principle

Conventional Antibody Labeling Process

STEP 1：Ultrafiltration buffer exchange and concentration (optional)

STEP 2：Antibody activation

STEP 3：Labeling reaction

STEP 4：Quench/Blocking and preservation

Other Issues:

Refer to the FAQ section