SKU: 53722538169

Protein L Donor Beads

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Description

Protein L Donor BeadsProduct Specification Stability & Storage Store at 2 8C away from light; product shelf life is 12 months. Background Homogeneous Immuno Chemiluminescence Assay (HICA) is a homogeneous immunoassay method based on energy transfer between donor beads and acceptor beads in 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

Product Specification


Stability & Storage

Store at 2-8°C away from light; product shelf life is 12 months.

Background

Homogeneous Immuno Chemiluminescence Assay (HICA) is a homogeneous immunoassay method based on energy transfer between donor beads and acceptor beads in 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 features a simple operation process, requiring no washing, and offers fast results with high sensitivity. It is capable of detecting weak interactions.

Components

Specification

Fill Volume

250 μg

50 μL

5 mg

1 mL

25 mg

1 mL x 5


Protocol

[Required Reagents]

Name

Catalog Number

Protein L Donor Beads

UA086111

Streptavidin Acceptor Beads

UA086090

Universal Buffer 1

UA086113


 

 

[Detection Procedure for Reference]

Detection Procedure

Detection Procedure 1 (37℃Rapid Detection)

Detection Procedure 2 (Room Temperature Detection)

Step 1:

4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Protein L Donor Beads,Protect from Light/Green Light

4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Protein L Donor Beads,Protect from Light/Green Light

Incubation

37℃ Shaking Incubation 20 minutes,Protect from Light/Green Light

Room Temperature Incubation 60 minutes,Protect from Light/Green Light

Step 2:

Add 6μL Streptavidin Acceptor Beads,Protect from Light/Green Light

Add 6μL Streptavidin Acceptor Beads,Protect from Light/Green Light

Incubation

37℃ Shaking Incubation 10 minutes,Protect from Light/Green Light

Room Temperature Incubation 30 minutes,Protect from Light/Green Light

Readout

Instrument Reading

Instrument Reading


 

 

[Performance Validation]

Sample Preparation:

Use Universal Buffer 1 to pre-dilute Biotinylated Human IgG (Bio-hIgG) to 15μg/mL (100nM) as stock solution, then perform gradient dilution according to the following scheme:

ID

Final Concentration (nM)

Universal Buffer 1

Volume (μL)

High Concentration Addition

Volume (μL)

C12

1.0E+01

210

90μL Stock Solution

C11

3.0E+00

210

90μL C12

C10

1.0E+00

180

90μL C11

C9

3.0E-01

210

90μL C10

C8

1.0E-01

180

90μL C9

C7

3.0E-02

210

90μL C8

C6

1.0E-02

180

90μL C7

C5

3.0E-03

210

90μL C6

C4

1.0E-03

180

90μL C5

C3

3.0E-04

210

90μL C4

C2

1.0E-04

180

90μL C3

C1

0

180

/


 

Detection Reagent Preparation:

Name

Preparation Concentration

Diluent

Protein L Donor Beads

25 μg/mL

Universal Buffer 1

Streptavidin Acceptor Beads

25 μg/mL

Universal Buffer 1


 

37℃ Incubation Mode Detection Results:

Max Signal: 450369

Min Signal: 616

EC50= 0.082 nM

Room Temperature Incubation Mode Detection Results:

Max Signal: 191706

Min Signal: 279

EC50= 0.059 nM

Guidelines

1. This experiment is light-sensitive; ensure all operations are performed in the dark. It is recommended to conduct preparation, sample addition, and incubation steps under green light (illuminance below 100 LUX). 2. This product is compatible with multi-functional microplate readers equipped with an Alpha detection module. 3. Vortex thoroughly before use. Alternatively, briefly centrifuge (2000×g, 5–10 seconds) to ensure complete retrieval. 4. It is recommended to use the company's accompanying dilution buffer for reagent preparation and sample dilution. If additional components are required, they can be directly added to this dilution buffer. 5. To ensure comparability of experimental data across different batches, strictly control incubation temperature and duration. 6. Avoid generating bubbles during sample addition.
Shipping Notes
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Exchange/Return Notes
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SKU: 53722538169

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