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Anti-FITC Acceptor Beads

Anti-FITC Acceptor Beads

Catalog Number: UA086101 Brand: UA BIOSCIENCE
Price:
Regular price $675 USD
Regular price Sale price $675 USD
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Product Details

Product Specification


Stability & Storage

Store in a dark place at 2~8℃; product shelf life is 12 months.

Background

Homogeneous Immuno Chemiluminescence Assay (HICA) is a homogeneous immunoassay based on energy transfer between donor beads and acceptor beads at close proximity, resulting in luminescence.

Donor beads recognize Protein 1 (Tag1 label), while Acceptor beads recognize Protein 2 (Tag2 label). When Protein 1 binds to Protein 2, the distance between the beads becomes less than 200nm. Upon excitation at 680nm, the donor beads generate singlet oxygen, which diffuses to the acceptor beads. The acceptor beads then undergo a redox reaction, emitting light at 615nm. The signal intensity is directly proportional to the strength of the protein interaction.

This product offers a simple, wash-free operation process, with high speed and sensitivity, enabling the detection of weak interactions.

Components

Specification

Volume

250 μg

50 μL

5 mg

1 mL

25 mg

1 mL x 5


Protocol

[Required Reagents]

Name

Catalog No.

Anti-FITC Acceptor Beads UA086101
Streptavidin Donor Beads UA086104
Universal Buffer 1 UA086113


[Detection Protocol for Reference]

Protocol

Protocol 1 (Rapid detection at 37°C)

Protocol 2 (Room temperature detection)

Step 1:

4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads,Protect from light / Green light

4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads,Protect from light / Green light

Incubation

Shake incubate at 37°C for 20 minutes,Protect from light / Green light

Incubate at room temperature for 60 minutes,Protect from light / Green light

Step 2:

Add 6μL Donor Beads,Protect from light / Green light

Add 6μL Donor Beads,Protect from light / Green light

Incubation

Shake incubate at 37°C for 10 minutes,Protect from light / Green light

Incubate at room temperature for 30 minutes,Protect from light / Green light

Reading

Instrument reading

Instrument reading


[Performance Validation]

Sample Preparation:

Dilute FITC-BSA-Biotin to 6.6 μg/mL (100 nM) as stock solution using Universal Buffer 1, then perform serial dilutions according to the following scheme:

No.

Final Concentration (nM)

Universal Buffer 1

Volume (μL)

High Conc. Addition

Volume (μL)

C12

1.0E+01

210

90 μL Stock Solution

C11

3.0E+00

210

90 μL C12

C10

1.0E+00

180

90 μL C11

C9

3.0E-01

210

90 μL C10

C8

1.0E-01

180

90 μL C9

C7

3.0E-02

210

90 μL C8

C6

1.0E-02

180

90 μL C7

C5

3.0E-03

210

90 μL C6

C4

1.0E-03

180

90 μL C5

C3

3.0E-04

210

90 μL C4

C2

1.0E-04

180

90 μL C3

C1

0

180

/


Detection Reagent Preparation:

Name

Preparation Concentration

Diluent

Anti-FITC Acceptor Beads

25 μg/mL

Universal Buffer 1

Streptavidin Donor Beads

25 μg/mL

Universal Buffer 1


Results for 37°C Incubation Mode:

Maximum Signal: 2779107

Minimum Signal: 1906

EC50= 0.290 nM

Results for Room Temperature Incubation Mode:

Maximum Signal: 1095538

Minimum Signal: 754

EC50= 0.334 nM

Guidelines

1. This experiment is light-sensitive; ensure all procedures (preparation, loading, and incubation) are performed in a dark environment. Recommended to operate under green light (illuminance below 100 LUX). 2. This product is compatible with multi-functional microplate readers equipped with Alpha detection modules. 3. Vortex thoroughly before use. Alternatively, briefly centrifuge (2000×g, 5–10 seconds) to ensure complete sample retrieval. 4. It is recommended to use the accompanying dilution buffer for reagent preparation and sample dilution. If additional components are required, they can be directly added to this buffer. 5. To ensure comparability of experimental data across different batches, strictly control incubation temperature and duration. 6. Avoid bubble formation during sample loading.