Product Overview
EnkiLife PerCP labeling kit contains multiple components for labeling antibody proteins, such as the activated lyophilized PerCP fluorescent dye, labeling buffer, and other chemical reagents, enabling easy and reliable antibody labeling for use as fluorescent probes.
Product Features
- Activated 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.
- Site-specific conjugation:Optimized site-specific conjugation technology ensures uniform antibody labeling and improves efficiency.
- Rapid process:The simple procedure allows for efficient labeling completion in as fast as 3 - 4 hours.
- High dye brightness: The selected high-performance fluorescent dye boasts high quantum yield and brightness, ensuring high detection sensitivity.
- Dye stability:The chosen high-performance fluorescent dye is stable and suitable for various subsequent applications, such as fixed sample detection.
- User-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.
- Flexibility: In the large-scale kit, the dye is pre-portioned for consistent batch performance, facilitating multiple labeling uses.
- Comprehensive 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
PerCP is a complex molecular dye composed of peridinin, chlorophyll, and lipids bound to peridinin-chlorophyll a protein. The peridinin molecule absorbs light in the blue-green wavelength range (470 nm to 550 nm) with a maximum emission at 675 nm. PerCP has a large Stokes shift and is well excited at 488 nm, with its maximum emission peak at 675 nm. Its emission spectrum is relatively narrow, which can greatly reduce the impact of spillover when used in multicolor flow Cytometry. Although PerCP itself has relatively weak intensity, its brightness can be enhanced when conjugated with other small molecule dyes such as Cy5.5.
Activated Dye Properties
Basic Characteristics
Synonyms:PCP
Molecular Weight: ~ 38 kDa
Common Excitation Sources: 488 nm
Maximum Excitation Wavelength:482 nm
Maximum Emission Wavelength: 677 nm
Molar Extinction Coefficient (L/(mol・cm)): 406000
Purity:A478/A280 ≥ 4.0
Solubility:Soluble in aqueous solutions
Appearance:Brown solid
Kit Specifications
| Product Components | Component Content in Different Specifications | Storage Temperature | ||
|---|---|---|---|---|
| 40μg Antibody | 200μg Antibody | 1mg Antibody | ||
| Activated PerCP | 1 tube | 1 tube | 5 tubes | -20°C after opening, protected from light |
| Labeling Buffer L | 2 mL | 2 mL | 2 mL | 4 °C, away from light |
| Antibody Modification Reagents | 60 μL | 60 μL | 60 μL | -20 °C, away from light |
| Ultrafiltration Tube * ,50K MWCO | 1 set ** | 1 set ** | 1 set ** | RT |
| Blocking reagent | 300 μg | 300 μg | 300 μg | -20 °C, away from light |
| DMSO (dissolve blocking reagent) | 50 μL | 50 μL | 50 μL | -20 °C, away from light |
| Labeled protein storage solution | 200 μL | 1 mL | 5 mL | 2-8 °C |
| Recommended amount of labeled antibody | Each tube dyes for labeling 20-40μg antibody Recommended: 20μg antibody | Each tube dyes for labeling 50-200μg antibody Recommended: 100μg antibody | Each tube dyes for labeling 50-200μg antibody Recommended: 100μg antibody | 2-8 °C |
* * Ultrafiltration tube molecular weight cutoff: 50KDa
** 1 set = 1 ultrafiltration tube and collection tube
Labeling Principle
This kit utilizes the cysteine characteristics of the antibody and the free amino groups on the dye to covalently couple the antibody molecule with the dye using a directed docking coupling technique.
Standard Labeling Procedure
- STEP 1：Ultrafiltration buffer exchange and concentration (optional)
- STEP 2：Antibody activation
- STEP 3：Labeling reaction
- STEP 4：Quench/Blocking and preservation
Data of PerCP labeling kit application
Flow Cytometry Analysis
FC experiment of mouse lymphocyte using antibodies preparaed with labeling kits and from the corresponding internationally brand company.
- Sample: mouse spleen lymphocyte cells from C57BL/6 mice
- Antibody:
Red histogram:0.05ug PerCP-anti Mouse CD4(GK1.5) from B company；
Gray histogram: 0.05ug PerCP-anti Mouse CD4(GK1.5) preparaed with PerCP labeling kits (RE80027) from EnkiLife； - Instrument: Agilent ACEA NovoCyte™3130
Conclusion: The antibody staining showed clear signals with a expectant signal-to-noise ratio. RE80027 kit is suitable for labeling anti-mouse CD4 antibody.
Flow Cytometry Analysis
Using the PerCP labeling kit(SKU:RE80027) and FITC labeling kit(SKU:RE80001p), label the anti-Mouse CD4 (GK1.5) and anti-Mouse CD3 antibody. After isolating the mouse spleen cells and preparing the cell suspension, red blood cell lysis, 10^6 cells per test, perform flow cytometry detection.
- Antibody:0.05ug PerCP-anti Mouse CD4(GK1.5) and 0.2ug FITC-anti mouse CD3 preparaed with two kinds of labeling kits from EnkiLife；
Conclusion: The antibodies preparaed with PerCP labeling kit and FITC labeling kit staining showed clear signals with a expectant signal-to-noise ratio.
