Time-resolved fluorescent microspheres - Particle size: 300nm; Surface functional group: - COOH; 10mg/ml in H2O

Cat. No.: T1521568

GRADE & PURITY Particle size: 300nm; Surface functional group: - COOH; 10mg/ml in H2O

Storage

Store at 2-8°C,Protected from light

Shipped In

Wet ice

Panoramica

This product is polystyrene microspheres doped with rare earth element europium (Eu). It has an excitation wavelength of 300–500 nm (365 nm is commonly used), and its emitted fluorescence features a large Stokes shift, long fluorescence lifetime and strong detection signals. Compared with conventional fluorescent materials, this product delivers high fluorescence signal intensity, making it more suitable for the development and mass production of high-sensitivity quantitative lateral flow assays.

Usage

I. One-Step Labeling Process

1. Remove the microsphere stock solution and disperse it using ultrasound.

2. Antibody coupling process.

Using a 200 μl volume for labeling as an example: Add 20 μl of (10 mg/ml) time-resolved fluorescent microspheres to 180 μl of 0.01 M PB (pH 6.2). After shaking to mix, add the antibody to a final concentration of 150 μg/ml, followed by 10 μl of EDC (0.1 mg/ml). Mix well and incubate with shaking for 2 hours. (Note: Dissolve EDC in 0.05 M MES buffer (pH 5.5); the labeling buffer should be adjusted appropriately; the buffers mentioned in the manual are for reference only)

3. Blocking: Add an appropriate amount of blocking solution and incubate with shaking at room temperature for 0.5–1 hour.

Add 100 μl of a 5% BSA aqueous solution and mix vertically at room temperature (blocking) for 0.5–1 hour.

4. Resuspension: Centrifuge, remove the supernatant, then add an appropriate amount of resuspension buffer to resuspend to the desired concentration; the sample is now ready for use.

Centrifuge at 10,000 rpm for 10 minutes, remove the supernatant, add 500 μl of resuspension buffer to adjust the concentration to 0.4 mg/ml, disperse by sonication, and store at 4°C;

II. Two-Step Labeling Procedure

1、Take out the microsphere stock solution and subject it to ultrasonic dispersion.

2、 Microsphere activation: Add EDC and NHS activation solution and react for 0.5~1h.

Taking the labeling of a 200ul volume as an example: Take 20ul (10mg/ml) of time-resolved fluorescent microspheres and add them to 180ul of 0.05M MES pH5.5 buffer containing 0.05% TW20. After shaking and mixing, add 2ul NHS (75 mg/ml) and 2ul EDC (50 mg/ml), and mix and shake for a reaction time of 0.5h. (Note: Both EDC and NHS are dissolved in 0.05M MES pH5.5 buffer.)

3、Resuspension: Centrifuge, remove the supernatant, and then resuspend with an appropriate buffer solution, followed by ultrasonic dispersion.

Centrifugation conditions: centrifuge at 10,000 rpm for 10 minutes, remove the supernatant, add 200 μl of 0.05M MES5.5 for resuspension, and then perform ultrasonic dispersion. (Centrifugation conditions should be adjusted appropriately according to changes in reaction volume, and the labeling buffer should be adjusted as needed.)

4、Conjugation: Add an appropriate amount of antibodies, antigens, nucleic acids, etc. to the above solution, and shake the reaction for 1-2 hours. If aggregation occurs during the reaction, perform ultrasonic dispersion.

Add 150 μg/ml antibody and conjugate for 1 hour. (If independent labeling is required, use a DNP-BSA labeling concentration of 150 μg/ml and react for another 1 hour.)

5、Blocking: Add an appropriate amount of blocking solution and shake at room temperature for 0.5~1h.

Add 100ul of 5% BSA in Tris solution (0.1M, pH 8.0) and mix vertically at room temperature for 0.5h (blocking).

6、Resuspending: Centrifuge, remove the supernatant, and add an appropriate amount of resuspension solution to resuspend to a quantitative concentration for use.

Centrifuge at 10000prm for 10min, remove the supernatant, add 500ul of resuspension solution to dilute to 0.4mg/ml, ultrasonicate for dispersion, and store at 4℃;

Precautions

1.Inappropriate use of buffer solution may lead to aggregation of markers. It is recommended to add ultrasonic operation during the marking process to disperse the microspheres.

2.Centrifugation conditions should be adjusted accordingly with changes in centrifuge volume.

3.Fluorescent microspheres need to be ultrasonically dispersed before use.

4.Coupling efficiency is affected by pH, temperature, molecular weight, and salt concentration. It is recommended to optimize the reaction conditions. 

5. Waste should be disposed of as hazardous waste.

Product Advantage

1.Surface functionalization: carboxyl groups；

2.Better fluorescence stability；

3.Fluorescent microspheres have uniform particle size, excellent monodispersity and good stability,The PDI index is all ≤ 0.3；

4.Higher detection sensitivity；

5.Suitable for two major technical platforms: quantitative immunochromatographic detection and microplate ultrasensitive detection.

Specifications

Specifiche e purezza

Particle size: 300nm; Surface functional group: - COOH; 10mg/ml in H2O

Condizioni di conservazione di stoccaggio

Store at 2-8°C,Protected from light

Spedito in

Wet ice

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.