CAPNS1/pACpET

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	CAPN1/pET or CAPN1:C115S/pET was co‐transformed into E. coli strain SoluBL21 (Genlantis) with CAPNS1/pACpET or CAPNS1ΔGR/pACpET, and the transformants were stored as frozen glycerol stocks at −80 °C. For protein expression and purification, the frozen transformants were precultured at 27 °C overnight, in 50 mL LB medium containing 100 μg/mL ampicillin and 50 μg/mL kanamycin (LB+Amp+Kn), and the cultures were then diluted into 1 L LB+Amp+Kn, and grown at 27 °C with vigorous shaking until A600 reached 0.4–0.5. The recombinant protein expression was induced by adding 1 mm isopropyl thiogalactoside (IPTG), followed by a 20‐h incubation at 24 °C, with vigorous shaking.

Protocol tips

CAPN1/pET or CAPN1:C115S/pET was co‐transformed into E. coli strain SoluBL21 (Genlantis) with CAPNS1/pACpET or CAPNS1ΔGR/pACpET, and the transformants were stored as frozen glycerol stocks at −80 °C. For protein expression and purification, the frozen transformants were precultured at 27 °C overnight, in 50 mL LB medium containing 100 μg/mL ampicillin and 50 μg/mL kanamycin (LB+Amp+Kn), and the cultures were then diluted into 1 L LB+Amp+Kn, and grown at 27 °C with vigorous shaking until A600 reached 0.4–0.5. The recombinant protein expression was induced by adding 1 mm isopropyl thiogalactoside (IPTG), followed by a 20‐h incubation at 24 °C, with vigorous shaking.

Publication protocol

CAPN1/pET or CAPN1:C115S/pET was co‐transformed into E. coli strain SoluBL21 (Genlantis) with CAPNS1/pACpET or CAPNS1ΔGR/pACpET, and the transformants were stored as frozen glycerol stocks at −80 °C. For protein expression and purification, the frozen transformants were precultured at 27 °C overnight, in 50 mL LB medium containing 100 μg/mL ampicillin and 50 μg/mL kanamycin (LB+Amp+Kn), and the cultures were then diluted into 1 L LB+Amp+Kn, and grown at 27 °C with vigorous shaking until A600 reached 0.4–0.5. The recombinant protein expression was induced by adding 1 mm isopropyl thiogalactoside (IPTG), followed by a 20‐h incubation at 24 °C, with vigorous shaking.

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Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Prokaryotic cells - E. coli rh μ‐calpain using CAPNS1/pACpET from Shoji Hata,Calpain Project, Department of Advanced Science for B.

Manufacturer protocol

Download the product protocol from Shoji Hata,Calpain Project, Department of Advanced Science for B for CAPNS1/pACpET below.

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Check out videos that might be relevant for performing Protein Expression Prokaryotic cells - E. coli rh μ‐calpain using CAPNS1/pACpET from Shoji Hata,Calpain Project, Department of Advanced Science for B. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.