Synthesis and application of secondary antibody taking graphene oxide as carrier
A graphene and carrier technology, applied in the synthesis and application of secondary antibodies, can solve the problems of difficult storage and transportation, high production cost of biological secondary antibodies, poor stability, etc., to shorten the experiment time, reduce transportation and storage costs , the effect of stability improvement
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Embodiment 1
[0038] Disperse 5-15 mg graphene oxide (GO) in 25 mL 2-morpholineethanesulfonic acid (MES) buffer (pH=6), sonicate vigorously for 2-3 h, add EDC (15 mg) and NHS (30 mg) at room temperature with gentle shaking for 12 h for activation. The activated GO was washed by centrifugation for 3 times, dispersed in PBS, added 10-25 mg of hexamethylenediamine, shaken at room temperature for 12 h, washed by centrifugation, and then dispersed in absolute ethanol (1% sodium cyanoborohydride). Add 10-25 mg of 2,6-difluoro-4-formylphenylboronic acid, shake for 6-12 h, and then add 100-200 μL of 1% bovine serum albumin (BSA) solution to passivate for 2 h to block the remaining active site. Add 100-250 μL of 5-40 mM phenolphthalein / DMSO solution, shake at room temperature for 6-12 h, centrifuge and wash 3 times, disperse in 5 mL 20 mMPBS (pH 7.4) and store until use.
Embodiment 2
[0040] Disperse 5-15 mg graphene oxide (GO) in 25 mL 2-morpholineethanesulfonic acid (MES) buffer (pH=6), sonicate vigorously for 2-3 h, add EDC (15 mg) and NHS (30 mg) at room temperature with gentle shaking for 12 h for activation. The activated GO was washed by centrifugation for 3 times, dispersed in PBS, added 10-25 mg of polyoxyethylene diamine (MW600) and shaken at room temperature for 12 h, washed by centrifugation, and then dispersed in absolute ethanol (1% cyanoboron Sodium hydride), add 10-25 mg of 2,6-difluoro-4-formylphenylboronic acid, shake for 6-12 h, then add 100-200 μL 1% bovine serum albumin (BSA) solution to passivate 2 h to block the remaining active sites. Add 100-250 μL of 5-40 mM phenolphthalein / DMSO solution, shake at room temperature for 6-12 h, centrifuge and wash 3 times, disperse in 5 mL of 20 mM PBS (pH7.4) and store until use.
Embodiment 3
[0042] Disperse 5-15 mg graphene oxide (GO) in 25 mL 2-morpholineethanesulfonic acid (MES) buffer (pH=6), sonicate vigorously for 2-3 h, add EDC (15 mg) and NHS (30 mg) at room temperature with gentle shaking for 12 h for activation. The activated GO was washed by centrifugation for 3 times, dispersed in PBS, added 10-25 mg of polyoxyethylene diamine (MW600) and shaken at room temperature for 12 h, washed by centrifugation, and then dispersed in absolute ethanol (1% cyanoboron Sodium hydride), add 10-25 mg of 2,4-difluoro-3-formylphenylboronic acid, shake for 6-12 h, then add 100-200 μL 1% bovine serum albumin (BSA) solution to passivate 2 h to block the remaining active sites. Add 10-100 μL of 5-30 mM thymolphthalein / DMSO solution, shake at room temperature for 6-12 h, centrifuge and wash 3 times, then disperse in 5 mL of 20 mM PBS (pH 7.4) and store until use.
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