The EnkiLife Cyanine5 labeling kit includes multiple components for labeling antibody proteins, such as high - performance fluorescent dye, ultrafiltration centrifuge tubes for purification, and labeling buffers. It enables easy and reliable labeling of antibodies and proteins.
Features
Fast: Labeling takes only about 30 minutes, balancing labeling effectiveness and efficiency.
Convenient: Each dye reagent is pre - optimized for corresponding antibody amounts, eliminating complex calculations. The solid dye form ensures batch stability, and following the steps yields good results.
Flexible: In the large - scale kit, the dye is pre - portioned for consistent batch performance, making it convenient for multiple labeling uses.
Outstanding labeling performance: The optimized labeling buffer ensures relatively fixed dye labeling sites, enhancing the homogeneity of labeled antibodies.
Amine - specific labeling: The activating group on the fluorescent dye efficiently labels antibodies and other purified proteins at primary amines.
Labeled proteins: This kit is mainly designed for IgG antibody labeling. Proteins similar in size and properties to IgG antibodies can also be labeled.
Fluorescent dye Introduction
Classic cyanine dyes, generally used for multicolor immunofluorescence, flow cytometry, or as tandem dyes.
Properties of Cyanine5
Similar Dyes: Alexa Fluor647,DyLight649,ATTO647
Excitation Wavelength: 646nm
Emission Wavelength: 662nm
Molar Extinction Coefficient (L/(mol・cm)): 271000
Purity: > 95%
Solubility: Soluble in organic solvents like DMSO and DMF
Chemical Structure:
Cyanine5 Spectrum
Kit Specifications
Product composition | Content of components in different specifications | Storage temperature | ||
40 μg antibody | 200 μg antibody | 2 mg antibody | ||
Cyanine5 | 1 tube | 5 tubes | 5 tubes | -20 °C after opening, away from light |
Ultrafiltration Tube* ,50K MWCO | 1 set ** | 1 set ** | 1 set ** | RT |
Labeling Buffer S | 10 mL | 10 mL | 10 mL | 2-8 °C |
1× PBS (pH 7.4) | 10 mL | 10 mL | 10 mL | 2-8 °C |
DMF | 100 μL | 100 μL | 100 μL | 2-8 °C, away from light |
Labeled protein storage solution | 200 μL | 1 mL | 5 mL | 2-8 °C |
Recommended amount of labeled antibody | Each tube is labeled with 20-40 μg, and the best is 20 μg | Each tube is labeled with 20-40 μg, and the best is 20 μg | Each tube is labeled with 100-400 μg, and the best is 200 μg | |
Application Examples of Cyanine5
Labeled antibodies are used as immunofluorescent probes.
Labeled primary antibodies are used in flow cytometry.
Labeled DNA/RNA probes are used for fluorescence in situ hybridization (FISH).
Labeled proteins are routinely used in difference gel electrophoresis (DIGE) (protein chips).
Labeled nanoparticles or antibodies are used to monitor the distribution of targets in living tissues in real-time (in vivo and small animal imaging).
Labeled PD-L1 antibodies are used to track changes in the tumor immune microenvironment.
Labeled antibodies are used with Cy3 for fluorescence resonance energy transfer (FRET) to validate protein-protein/DNA-protein interactions.
Cy5-labeled secondary antibodies replace traditional chemiluminescence, allowing direct quantification of target band fluorescence for high-sensitivity Western blotting of low-abundance proteins.
Labeled antibodies are combined with PE dyes to form tandem dyes, creating new fluorescent dyes (based on FRET principles).
Cy5-labeled microspheres are coupled with capture antibodies. Flow cytometry is used to distinguish target molecules, enabling multiplex detection (liquid phase chips, multi-factor detection).
Cyanine5 - Antibody Labeling Principle
Within a certain pH range, the activated group of the fluorescent dye specifically reacts with primary amino groups on antibody proteins to form stable amide bonds, thereby conjugating the dye to the antibody protein.
Conventional Antibody Protein Labeling Process
Step 1：Ultrafiltration buffer exchange and concentration
Step 2：Labeling reaction
Step 3：Purification
Step 4：DOL calculation and collection preservation
