Myo detection kit based on bimolecular fluorescence complementation technology and preparation and use method thereof
A detection kit, bimolecular technology, applied in biological testing, fluorescence/phosphorescence, analytical materials, etc., can solve problems such as reducing detection repeatability
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] The anti-Myo antibody is coupled to the N-terminal fragment of fluorescent protein, taking the yellow fluorescent protein (YFP) 1-154 amino acid fragment YFPN as an example, the specific implementation process is as follows:
[0027] 1) Add 0.1mg of YFPN protein into a centrifuge tube, and prepare with 0.05mol / L pH9.5 carbonate buffer (CB) to dilute the YFPN protein to 1mg / ml.
[0028] 2) Add glutaraldehyde at a final concentration of 1.25% in a fume hood.
[0029] 3) Reaction in a water bath at 37°C for 2 hours.
[0030] 4) Dialyze with desalting column Sephadex G-25 or 0.05mol / L pH9.5 CB to remove excess glutaraldehyde.
[0031] 5) Take 0.1 mg anti-Myo antibody, prepare 1 mg / ml antibody with 0.05 mol / L pH9.5 CB, and mix activated YFPN protein and antibody.
[0032] 6) React overnight at 4°C.
[0033] 7) Blocking: add 50 μl of 0.2 mol / L lysine solution, and block for 2 hours at room temperature to block residual aldehyde groups and terminate the reaction.
[0034] ...
Embodiment 2
[0037] The anti-Myo antibody is coupled to the fluorescent protein C-terminal fragment, taking the yellow fluorescent protein (YFP) 155-238 amino acid fragment YFPC as an example, the specific implementation process is as follows:
[0038] 1) Add 0.1mg of YFPC protein into a centrifuge tube and prepare with 0.05mol / L pH9.5 carbonate buffer (CB) to dilute the YFPC protein to 1mg / ml.
[0039] 2) Add glutaraldehyde at a final concentration of 1.25% in a fume hood.
[0040] 3) Reaction in a water bath at 37°C for 2 hours.
[0041] 4) Dialyze with desalting column Sephadex G-25 or 0.05mol / L pH9.5 CB to remove excess glutaraldehyde.
[0042] 5) Take 0.1 mg anti-Myo antibody, prepare 1 mg / ml antibody with 0.05 mol / L pH9.5 CB, and mix activated YFPC protein and antibody.
[0043] 6) React overnight at 4°C.
[0044] 7) Blocking: add 50 μl of 0.2 mol / L lysine solution, and block for 2 hours at room temperature to block residual aldehyde groups and terminate the reaction.
[0045] 8)...
Embodiment 3
[0048] The main components of the kit:
[0049] 1) N-terminal fragment of fluorescent protein coupled with anti-Myo antibody;
[0050] 2) Anti-Myo antibody-coupled fluorescent protein C-terminal fragment.
PUM
| Property | Measurement | Unit |
|---|---|---|
| Analytical sensitivity | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More