Human IgG Donor Beads

Catalog Number: UA086112 Brand: UA BIOSCIENCE

Price:

$144 USD

Size: 250μg 5mg 25mg

Quantity:

For shipping services or bulk orders, you may request a quotation.

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Product Details

Product Specification

Stability & Storage

Store in a dark place at 2-8°C; 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), and acceptor beads recognize protein 2 (Tag2 label). When protein 1 binds to protein 2, the distance between the beads is less than 200 nm. Upon excitation at 680 nm, donor beads generate singlet oxygen, which diffuses to acceptor beads. The acceptor beads then undergo a redox reaction, emitting light at 615 nm. The signal intensity is proportional to the strength of protein interaction.

This product features a simple operation process, no washing steps, fast speed, high sensitivity, and the ability to detect weak interactions.

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Protocol

[Required Reagents]

Name	Catalog Number
Human IgG Donor Beads	UA086112
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 Human IgG Donor Beads，Avoid Light / Green Light	4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Human IgG Donor Beads，Avoid Light / Green Light
Incubation	37℃ Shaking Incubation 20 minutes，Avoid Light / Green Light	Room Temperature Incubation 60 minutes,，Avoid Light / Green Light
Step 2:	Add 6μL Streptavidin Acceptor Beads，Avoid Light / Green Light	Add 6μL Streptavidin Acceptor Beads，Avoid Light / Green Light
Incubation	37℃ Shaking Incubation 10 minutes，Avoid Light / Green Light	Room Temperature Incubation 30 minutes，Avoid Light / Green Light
Reading	Instrument Reading	Instrument Reading

[Performance Validation]

•Sample Preparation:

Use Universal Buffer 1 to pre-dilute biotinylated rabbit IgG (Bio-rIgG) to 15μg/mL (100nM) as a 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
Human IgG Donor Beads	25 μg/mL	Universal Buffer 1
Streptavidin Acceptor Beads	25 μg/mL	Universal Buffer 1

•37℃ Incubation Mode Results:

Maximum Signal: 390419

Minimum Signal: 409

EC50= 0.076 nM

•Room Temperature Incubation Mode Results:

Maximum Signal: 194000

Minimum Signal: 181

EC50= 0.043 nM

Guidelines

1. This experiment is light-sensitive; ensure all operations are performed in the dark. It is recommended to carry out 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 recommended to use the company’s accompanying diluent for reagent preparation and sample dilution. If additional components are required, they can be directly added to this diluent. 5. To ensure comparability of experimental data across different batches, strictly control incubation temperature and time. 6. Avoid generating bubbles during sample loading.

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Human IgG Donor Beads

Human IgG Donor Beads