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Antipain dihydrochloride

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Antipain Dihydrochloride is a protease inhibitor used in biochemical research applications. It inhibits the activity of serine and cysteine proteases. The product is available in powder form.

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Lab products found in correlation

Aprotinin, by GoldBio (2 mentions) Leupeptin, by GoldBio (2 mentions)

2 protocols using antipain dihydrochloride

1

Assembly of SDS-resistant SNARE Complex

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The SNARE motifs were mixed in equimolar ratios in the following order: synaptobrevin-2 (29–93), SNAP-25A(141–203), SNAP-25A(11–82) and syntaxin-1A (191–253), in the presence of 1 M NaCl. The mixture contained the following protease inhibitors (protease inhibitor cocktail A): Antipain Dihydrochloride 0.016 mg/ml (Thermo Fischer Scientific: 50488492); Leupeptin 0.33 mg/ml (Gold Bio: L01025); Aprotinin 0.08 mg/ml (Gold Bio: A655100). The assembly reaction was incubated at room temperature overnight while rotating. The SNARE motifs that did not incorporate into complex were removed by concentration-dilution at room temperature using 30 kDa molecular weight cutoff (MWCO) Amicon centrifugation filters. The almost quantitative formation of SDS-resistant SNARE complex was verified by SDS-PAGE and Coomassie blue staining.
Voleti R., Bali S., Guerrero J., Smothers J., Springhower C., Acosta G.A., Brewer K., Albericio F, & Rizo J. (2021). Evaluation of the tert-butyl group as a probe for NMR studies of macromolecular complexes. Journal of biomolecular NMR, 75(8-9), 347-363.
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2

Assembling SNARE Protein Complex

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The SNARE motifs were mixed in the equimolar ratio in following order: synaptobrevin-2 (29–93), SNAP-25A(141–203), SNAP-25A(11–82) and syntaxin-1A (191–253), in the presence of 1 M NaCl. The mixture contained the following protease inhibitors (protease inhibitor cocktail A): Antipain Dihydrochloride 0.016 mg/ml (Thermo Fischer Scientific: 50488492); Leupeptin 0.33 mg/ml (Gold Bio: L01025); Aprotinin 0.08 mg/ml (Gold Bio: A655100). The assembly reaction was incubated in room temperature overnight while rotating. The SNARE motifs that did not incorporate into complex were removed by concentration-dilution at room temperature using 30 kDa molecular weight cutoff (MWCO) Amicon centrifugation filters.
Voleti R., Jaczynska K, & Rizo J. (2020). Ca2+-dependent release of synaptotagmin-1 from the SNARE complex on phosphatidylinositol 4,5-bisphosphate-containing membranes. eLife, 9, e57154.
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