Methods and reagents for the detection of biomolecules using luminescence
A technology of biomolecules and biotin, which is applied in the field of competitive binding assays, can solve problems such as difficult to change, rare materials, expensive, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0301] Embodiment 1: Use the streptavidin fluorescence enhancement of AQC-CTC to carry out biotinylated oligonucleotide detection Measurement
[0302] As shown in Table 1, prepare three different solutions containing 15 μl of AQC-CTC (150 mg / l), 55 μL of streptavidin solution (0.5 mg / ml in water) and increasing amounts of oligonucleotide-biotin solution (1 nM in TE buffer). After 1 hour of incubation, the fluorescence of each sample was measured.
[0303] table 3
[0304]
[0305] figure 1 The emission spectra (measured in a quartz cuvette (quartzcell) with a light path of 3×3 mm and a volume of 45 μl with a CaryEclipse Varian fluorescence spectrophotometer) of different solutions: (A ) AQC-CTC with streptavidin; (B) AQC-CTC with streptavidin and 0.1 mmol oligonucleotide-biotin; and (C) AQC-CTC with streptavidin and 0.5 mmol Oligonucleotides - Biotin. The spectrum shows an emission maximum at 600 nm which decreases with the addition of oligonucleotide-biotin and wh...
Embodiment 2
[0306] Example 2: Biotin with Streptavidin / HABA Fluorescence Enhancement / Quenching of AQC-CTC Chemical oligonucleotide detection
[0307] As shown in Table 2, two solutions were prepared as follows: 0.25 μl of HABA solution (10 mM) and 0.5 μl of AQC-CTC solution (about 150 mg / l) were added to a specific volume of streptavidin solution (0.5 mg / ml). Then, 0.5 μl of oligonucleotide-biotin solution (1 nM) was added to solution B and both samples were incubated for 30 minutes.
[0308] Table 4
[0309]
[0310] figure 2 The emission spectra (measured in a quartz cuvette with a light path of 3 × 3 mm and a volume of 45 μl with a CaryEclipse Varian fluorescence spectrophotometer) of different solutions: (A) AQC- CTC with HABA and streptavidin; and (B) AQC-CTC with HABA, streptavidin and 0.5 μl oligonucleotide-biotin. Such as figure 2 As shown in , the addition of oligo-biotin resulted in a decrease in the maximum emission intensity, which was attributed to the int...
Embodiment 3
[0311] Example 3. Method for coupling protein molecules to nanovesicles
[0312] Freshly prepared aqueous solution of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) was added to the solution of nanovesicles in pH 9 borate buffer. After 5 minutes of moderate stirring, the protein solution in pH 9 borate buffer was added. Stirring was continued for 2 hours and the final protein-nanovesicle conjugate was purified by centrifugal ultrafiltration.
PUM
| Property | Measurement | Unit |
|---|---|---|
| thickness | aaaaa | aaaaa |
| diameter | aaaaa | aaaaa |
| affinity | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More