pET-21b(+)-mTNFα

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The selected plasmid was then transformed into E. coli BL21 (DE3) cells, which were cultured in 3 mL of LB medium containing 100 μg/mL ampicillin in logarithmic phase (OD 600 of 0.6). Isopropyl-beta-D-thiogalactopyranoside (IPTG) was then added in a concentration of 0.4 μg/mL to induce protein expression.	After 18 hours of incubation at 37 °C, the cells were pelleted in a microfuge at 13400 (RCF) for 1 (min). SDS-polyacrylamide gel electrophoresis (SDS-PAGE) was performed on both the cell pellet and supernatant to detect protein expression.

Protocol tips
The selected plasmid was then transformed into E. coli BL21 (DE3) cells, which were cultured in 3 mL of LB medium containing 100 μg/mL ampicillin in logarithmic phase (OD 600 of 0.6). Isopropyl-beta-D-thiogalactopyranoside (IPTG) was then added in a concentration of 0.4 μg/mL to induce protein expression.
Downstream tips
After 18 hours of incubation at 37 °C, the cells were pelleted in a microfuge at 13400 (RCF) for 1 (min). SDS-polyacrylamide gel electrophoresis (SDS-PAGE) was performed on both the cell pellet and supernatant to detect protein expression.

Publication protocol

The selected plasmid was then transformed into E. coli BL21 (DE3) cells, which were cultured in 3 mL of LB medium containing 100 μg/mL ampicillin in logarithmic phase (OD 600 of 0.6). Isopropyl-beta-D-thiogalactopyranoside (IPTG) was then added in a concentration of 0.4 μg/mL to induce protein expression. After 18 hours of incubation at 37 °C, the cells were pelleted in a microfuge at 13400 (RCF) for 1 (min). SDS-polyacrylamide gel electrophoresis (SDS-PAGE) was performed on both the cell pellet and supernatant to detect protein expression.

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