CAPNS1ΔGR/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 CAPNS1ΔGR/pACpET from Shoji Hata,Calpain Project, Department of Advanced Science for B.

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Download the product protocol from Shoji Hata,Calpain Project, Department of Advanced Science for B for CAPNS1ΔGR/pACpET below.

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