SKU: 5537051972

Anti-His Tag Acceptor Beads

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Description

Anti-His Tag Acceptor BeadsProduct Specification Stability & Storage Store away from light at 2 8; 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 for 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

Product Specification


Stability & Storage

Store away from light at 2-8℃; 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 for 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, 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 protein interaction.

This product features a simple operation process, no washing steps, fast speed, and high sensitivity, enabling the detection of weak binding.

Components

Specification

Volume

250 μg

50 μL

5 mg

1 mL

25 mg

1 mL x 5


Protocol

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【Required Reagents】

Name

Catalog Number

Anti-His Tag Acceptor Beads UA086100
Streptavidin Donor Beads UA086104
Universal Buffer 1 UA086113


 

【Detection Procedure (For Reference)】

Detection Steps

Procedure 1 (37°C Rapid Detection)

Procedure 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

​37°C shaking incubation for 20 minutes,Protect from light/green light

​Room temperature incubation 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

​37°C shaking incubation for 10 minutes,Protect from light/green light

​Room temperature incubation for 30 minutes,Protect from light/green light

Reading

Instrument reading

Instrument reading


 

【Performance Validation】

Sample Preparation:

Biotinylated 6xHis (Bio-HH-6) was pre-diluted to 11μg/mL (10μM) using Universal Buffer 1 as the stock solution, followed by serial dilution as below:

ID

Final Concentration (nM)

Universal Buffer 1 Volume (μL)

High Concentration Addition Volume (μL)

C12

1.0E+03

210

90μL stock

C11

3.0E+02

210

90μL C12

C10

1.0E+02

180

90μL C11

C9

3.0E-01

210

90μL C10

C8

1.0E-01

180

90μL C9

C7

3.0E-00

210

90μL C8

C6

1.0E-00

180

90μL C7

C5

3.0E-01

210

90μL C6

C4

1.0E-01

180

90μL C5

C3

3.0E-02

210

90μL C4

C2

1.0E-02

180

90μL C3

C1

0

180

/


 

Detection Reagent Preparation:

Name

Preparation Concentration

Diluent

Anti-His Tag Acceptor Beads

25 μg/mL

Universal Buffer 1

Streptavidin Donor Beads

25 μg/mL

Universal Buffer 1


 

 

37°C Incubation Mode Results:

 

Maximum signal: 222589

Minimum signal: 569

EC50= 3.919 nM

Room Temperature Incubation Mode Results:

Maximum signal: 119656

Minimum signal: 316

EC50= 3.656 nM

```

Guidelines


1. This experiment is light-sensitive. Perform all procedures 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. Alternatively, briefly centrifuge (2000×g, 5–10 seconds) to ensure complete sample retrieval.

4. It is recommended to use the accompanying dilution buffer provided by our company for reagent preparation and sample dilution. For special additive requirements, corresponding components 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: 5537051972

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