Common use of Conjugation Clause in Contracts

Conjugation. During conjugate preparation, the single cysteine mutants will form Au-S bonds with ▇▇▇▇▇ and the His tag mutant will strongly associate with the His-Tag. Each mutant was rehydrated with 1.42 mg/mL TCEP, to prevent disulfide bonds from forming,30 and were left to incubate for at least an hour. To make 5 nm conjugates, 400 μL of 2.40 nM 5 nm AuNPs were added to 30 μL of 50 mM free protein in a lo-bind Eppendorf tube and left to incubate overnight at 4°C to allow the conjugation to occur. The samples were centrifuged for 90 minutes at 13,200 rpm. The supernatant was removed, and the pellet was replenished with 0.005% Tween in 10 mM sodium phosphate buffer to wash the free protein from the pellet of DHFR-AuNP conjugates. After wash cycles, the pellet was stored in 0.005% Tween in 10 mM sodium phosphate buffer, and diluted accordingly. To make 15 nm conjugates, the same methods to make 5 nm conjugates were made except centrifuging speeds and times were changed to 8000 rpm and 35 minutes for 800 μL of 7.88 nM 15 nm AuNP. To make 30 nm conjugates, the same methods to make 5 nm conjugates were made except centrifuging speeds and times were changed to 8000 rpm and 10 minutes for 400 μL of

Conjugation. During conjugate preparation, the single cysteine mutants will form Au-S bonds with ▇▇▇▇▇ AuNPs and the His tag mutant will strongly associate with the His-Tag. Each mutant was rehydrated with 1.42 mg/mL TCEP, to prevent disulfide bonds from forming,30 and were left to incubate for at least an hour. To make 5 nm conjugates, 400 μL L of 2.40 nM 5 nm AuNPs were added to 30 μL L of 50 mM free protein in a lo-bind Eppendorf tube and left to incubate overnight at 4°C to allow the conjugation to occur. The samples were centrifuged for 90 minutes at 13,200 rpm. The supernatant was removed, and the pellet was replenished with 0.005% Tween in 10 mM sodium phosphate buffer to wash the free protein from the pellet of DHFR-AuNP conjugates. After wash cycles, the pellet was stored in 0.005% Tween in 10 mM sodium phosphate buffer, and diluted accordingly. To make 15 nm conjugates, the same methods to make 5 nm conjugates were made except centrifuging speeds and times were changed to 8000 rpm and 35 minutes for 800 μL L of 7.88 nM 15 nm AuNP. To make 30 nm conjugates, the same methods to make 5 nm conjugates were made except centrifuging speeds and times were changed to 8000 rpm and 10 minutes for 400 μL ofL of 2.40 nM 15 nm AuNP.