SKU: 49546527630

Mouse Monoclonal Antibody Screening Set (Flag-tag Antigen)

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Description

Mouse Monoclonal Antibody Screening Set (Flag-tag Antigen)Product Specification Stability & Storage Store at 2 8C away from light; product shelf life is 12 months. Background 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,

Product Specification


Stability & Storage

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

Background

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 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 high speed and sensitivity. It is capable of detecting weak interactions.

Components

Name

Main Components

3000T

Reagent A

Anti-Flag-tag antibody-crosslinked acceptor Beads

10mL

Reagent B

Biotin-labeled anti-mouse IgG antibody

10mL

Reagent D

Streptavidin Donor Beads

10mL


Protocol

[Required Reagents]

Name

Catalog Number

Universal Buffer 4

UA086115


 

 

[Detection Procedure for Reference]

Detection Process

Detection Process

Step 1:

Add Flag-tag antigen to Reagent A at a concentration of 1.5μg/mL

Step 2:

Mix the above Reagent A, Reagent B, and Reagent D uniformly at a volume ratio of 1:1:1,avoid light/green light

Step 3:

Add 10μL of test sample + 10μL of mixed reagent per test,avoid light/green light

Incubation

​37°C with shaking for 60 minutes,avoid light/green light

Reading

Instrument reading


 

[Sample and Reagent Preparation]

Pre-dilute mouse anti-IL1B antibody to 1000ng/mL (6.67nM) using DMEM+10%FBS as C8, then perform gradient dilution according to the following scheme:

ID

Concentration

(ng/mL)

Diluent Volume

(μL)

Antibody Volume

(μL)

C8

1000

/

/

C7

200

160

40μL C8

C6极>

40

160

40μL C7

C5

8

160

40μL C6

C4

1.6

160

40μL C5

C3

0.32

160

40μL C4

C2

0.064

160

40μL C3

C1

0

160

/


 

 

 

 

 

[Performance Validation]

Response curve:

Maximum signal: 1048745

Minimum signal: 280

EC50= 2.106 ng/mL (0.014 nM)

 

Limit of blank:

Repeat检测20次C1,计算信号均值及SD,用反应曲线计算 信号均值+2×SD对应的浓度值,即为空白限。

Limit of blank= 0.738 ng/mL (4.922 pM)

 

Repeatability:

Repeat testing of high and low concentration samples 10 times, calculate the concentration CV%.

Repeatability

Low concentration

High concentration

CV%

1.3%

3.3%


 

 

 

 

 

Specificity: Dilute the following proteins to 1μg/mL using DMEM+10%FBS and test the cross-reactivity rate.

Test substance

Cross-reactivity rate

Human anti-IL1B antibody

0.00%


 

 

 

Guidelines


1. This experiment is light-sensitive. Please protect it from light during operation. It is recommended to perform 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 Alpha detection modules.

3. To ensure comparability of experimental data across different batches, please strictly control incubation temperature and time.

4. Avoid generating bubbles during sample addition.

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SKU: 49546527630

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Easy to handle, hard to crimp.
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This cable is easy to manage (bend, etc) because it has an aluminum core. But for that same reason it is harder to effectively crimp, even with a hydraulic crisper.
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