SKU: 12032573010

Anti-Human IgG Donor Beads

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Description

Anti-Human IgG Donor BeadsProduct Specification Stability & Storage Store at 28C 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 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

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 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 features a simple operation process, requires no washing, and offers fast results with high sensitivity. It is capable of detecting weak interactions.

Components

Specification

Volume

250 μg

50 μL

5 mg

1 mL

25 mg

1 mL x 5


Protocol

[Required Reagents]

Name

Cat. No.

Anti-Human IgG Donor Beads

UA086109

Streptavidin Acceptor Beads

UA086090

Universal Buffer 1

UA086113


[Assay Procedure for Reference]

Assay Procedure

Assay Procedure 1 (37°C Rapid Assay)

Assay Procedure 2 (Room Temperature Assay)

Step 1:

4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Anti-Human IgG Donor Beads, Protect from light / Green light

4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Anti-Human IgG Donor Beads, Protect from light / Green light

Incubation

Shake and 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 Streptavidin Acceptor Beads, Protect from light / Green light

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

Incubation

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

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

Readout

Instrument Readout

Instrument Readout


[Performance Validation]

Sample Preparation:

Pre-dilute biotinylated human IgG (Bio-hIgG) to 15 μg/mL (100 nM) using Universal Buffer 1 as the stock solution, then perform serial dilutions according to the following scheme:

No.

Final Concentration (nM)

Volume of Universal Buffer 1 (μL)

Volume of High Conc. Addition (μL)

C12

1.0E+01

210

90 μL 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

Working Concentration

Diluent

Anti-Human IgG Donor Beads

25 μg/mL

Universal Buffer 1

Streptavidin Acceptor Beads

25 μg/mL

Universal Buffer 1


37°C Incubation Mode Results:

Maximum Signal:47550

Minimum Signal:192

EC50= 0.067 nM

Results of Room Temperature Incubation Mode:

Maximum Signal:23402

Minimum Signal:92

EC50= 0.036 nM

Guidelines

1. This experiment is light-sensitive; ensure all procedures are performed under light-protected conditions. It is recommended to conduct preparation, sample loading, and incubation steps under green light (illuminance below 100 LUX). 2. This product is compatible with multifunctional microplate readers equipped with Alpha detection modules. 3. Vortex thoroughly before use, or briefly centrifuge (2000×g, 5–10 seconds) to ensure complete sample retrieval. 4. It is advised to use the accompanying dilution buffer from our company 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 generating bubbles during sample loading.
Shipping Notes
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Exchange/Return Notes
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SKU: 12032573010

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