ft4 in vitro diagnostic kit and method of use thereof
A kit, FT4 technology, which is applied in biological testing, analysis by chemical reaction of materials, and material testing products, can solve problems such as poor hapten coating, achieve stability and extend the validity period, eliminate environmental and Hazards to operators, effects of getting rid of half-life limitations
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Embodiment 1
[0053] Material:
[0054] Activation buffer: 0.1MMES, 0.5MNACl, pH6.0;
[0055] Carrier protein: BSA;
[0056] Target protein: small molecule hapten FT4;
[0057] NHS; Mercaptoethanol; Desalting column
[0058] Coupling the small molecule hapten FT4 with the carrier protein,
[0059] ① Equilibrate EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), NHS (N-hydroxysuccinimide) to room temperature.
[0060] ②Dissolve EDC with activation buffer, the final concentration is 50mg / mL, NHS is also dissolved with activation buffer, the final concentration is 50mg / mL, BSA (bovine serum albumin) is dissolved with activation buffer, the final concentration is 5mg / mL .
[0061] ③ After mixing 200uLEDC solution with 500uLBSA solution, add 200uL NHS solution, react at room temperature for 15 minutes, and then add 100uLEDC. React for another 15 minutes.
[0062] ④Add 1.4uL mercaptoethanol to terminate the EDC reaction and obtain the activated protein (EDC-BSA).
[0063] ...
Embodiment 2
[0087] Material:
[0088] Activation buffer: 0.1MMES, 0.5MNACl, pH6.0
[0089] Carrier protein: BSA;
[0090] Target protein: small molecule hapten FT4
[0091] NHS; Mercaptoethanol; Desalting column
[0092] Coupling the small molecule hapten FT4 with the carrier protein,
[0093] ① Equilibrate EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), NHS (N-hydroxysuccinimide) to room temperature.
[0094] ②Dissolve EDC with activation buffer, the final concentration is 45mg / mL, NHS is also dissolved with activation buffer, the final concentration is 55mg / mL, BSA (bovine serum albumin) is dissolved with activation buffer, the final concentration is 4.5mg / mL mL.
[0095] ③ After mixing 200uLEDC solution with 500uLBSA solution, add 200uL NHS solution, react at room temperature for 15 minutes, and then add 100uLEDC. React for another 15 minutes.
[0096] ④Add 1.4uL mercaptoethanol to terminate the EDC reaction and obtain the activated protein (EDC-BSA).
[0097...
Embodiment 3
[0119] Material:
[0120] Activation buffer: 0.1MMES, 0.5MNACl, pH6.0
[0121] Carrier protein: BSA;
[0122] Target protein: small molecule hapten FT4
[0123] NHS; Mercaptoethanol; Desalting column
[0124] The method of coupling the small molecule hapten FT4 and the carrier protein is as follows:
[0125] ① Equilibrate EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), NHS (N-hydroxysuccinimide) to room temperature.
[0126] ②Dissolve EDC with activation buffer, the final concentration is 55mg / mL, NHS is also dissolved with activation buffer, the final concentration is 45mg / mL, BSA (bovine serum albumin) is dissolved with activation buffer, the final concentration is 5.0mg / mL mL.
[0127]③ After mixing 200uLEDC solution with 500uLBSA solution, add 200uL NHS solution, react at room temperature for 15 minutes, and then add 100uLEDC. React for another 15 minutes.
[0128] ④Add 1.4uL mercaptoethanol to terminate the EDC reaction and obtain the activated p...
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