Mouse Monoclonal Antibody Screening Set (GST-tag Antigen)

The 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 200 nm. Upon excitation at 680 nm, the donor beads generate singlet oxygen, which diffuses to the Acceptor beads. The Acceptor beads then undergo a redox reaction, emitting light at 615 nm. 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 high speed and sensitivity, enabling the detection of weak interactions.

Name	Main Component	3000T
Reagent A	Anti-GST-tag antibody-crosslinked acceptor Beads	10mL
Reagent B	Biotin-labeled anti-mouse IgG antibody	10mL
Reagent D	Streptavidin Donor Beads	10mL

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[Required Reagents]

Name	Catalog No.
Universal Buffer 4	UA086115

[Detection Protocol for Reference]

Detection Step	Procedure
Step 1:	Add GST-tag antigen to Reagent A at 0.5μg/mL concentration
Step 2:	Mix Reagent A, B, and D uniformly at a 1:1:1 volume ratio, protect from light/green light
Step 3:	Add 10μL test sample + 10μL mixed reagent per test, protect from light/green light
Incubation	37°C shaking incubation for 60 minutes, protect from light/green light
Reading	Instrument reading

[Sample and Reagent Preparation]

•Pre-dilute M-1b antibody to 1000ng/mL (6.67nM) with Universal Buffer 4 as C8, then perform serial dilution as follows:

ID	Concentration (ng/mL)	Universal Buffer 4 Volume (μL)	High-Concentration Addition Volume (μL)
C8	1000	/	/
C7	200	160	40μL C8
C6	40	160	40μL C7
C5	8	160	40μL C6
C4	1.6	160	40μL C5
Name	Catalog No.
Universal Buffer 4	UA086115

[Procedure]

Step	Action
1	Add GST-tag antigen (0.5μg/mL) to Reagent A
2	Mix A:B:D (1:1:1), light-sensitive
3	Add 10μL sample + 10μL mixed reagent, light-sensitive
Incubation	37°C, 60min, light-sensitive
Reading	Instrument measurement

[Sample Prep]

ID	Conc.(ng/mL)	Buffer(μL)	Add(μL)
C8	1000	-	-
C7	200	160	40 C8

[Validation]

• Dose-response: Max=1,830,280, Min=943, EC50=126.5ng/mL (0.84nM)
• LoB: 0.099ng/mL (0.66pM)
• CV%: Low=2.2%, High=1.1%
• Specificity: Human IgG=0%, Rabbit IgG=0%

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1. This experiment is light-sensitive; ensure all operations are performed in the dark. It is recommended to conduct preparation, sample loading, and incubation under green light (illuminance below 100 LUX). 2. This product is compatible with multi-mode microplate readers equipped with Alpha detection modules. 3. To ensure comparability of experimental data across different batches, strictly control incubation temperature and time. 4. Avoid generating bubbles during sample loading.