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Merck

Purification of Recombinant Galectins Expressed in Bacteria.

STAR protocols (2020-12-31)
Cecilia Arahí Prato, Julieta Carabelli, Valentina Cattaneo, Oscar Campetella, María Virginia Tribulatti
要旨

Galectins are soluble lectins that participate in many physiological and pathological functions. Since they can act extracellularly, the use of the recombinant protein is a recurrent strategy for studying their biological functions. Here, we provide a general protocol for the production of Galectins and their isolated or chimeric domains. We take advantage of their lectin activity and the 6xHis-tag addition for purification, thus obtaining a highly pure and active Galectin to use in both in vitro and in vivo assays. For complete details on the use and execution of this protocol, please refer to Cattaneo et al. (2011), Tribulatti et al. (2012), and Prato et al. (2020).

材料
製品番号
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製品内容

Sigma-Aldrich
デオキシリボヌクレアーゼ I ウシ膵臓由来, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein
Sigma-Aldrich
リゾチーム ニワトリ卵白由来, powder or granules, ≥39,000 units/mg protein
Sigma-Aldrich
過硫酸アンモニウム, Molecular Biology, suitable for electrophoresis, ≥98%
Sigma-Aldrich
水酸化ナトリウム, BioXtra, ≥98% (acidimetric), pellets (anhydrous)
Sigma-Aldrich
Triton X-100 溶液, BioUltra, Molecular Biology, ~10% in H2O
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フェニルメチルスルホニルフルオリド, ≥98.5% (GC)
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Yeast Extract, for use in microbial growth medium
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ヨードアセトアミド, ≥99% (NMR), crystalline
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アジ化ナトリウム, BioUltra, ≥99.5% (T)
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D-ラクトース 一水和物, ≥99% (HPLC), BioUltra
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Trizma® 塩酸塩, BioUltra, Molecular Biology, ≥99.0% (AT)
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ドデシル硫酸ナトリウム, ACS reagent, ≥99.0%
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塩化ナトリウム, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
α-ラクトース-アガロース, saline suspension
Sigma-Aldrich
EZMix トリプトン, Pancreatic digest of casein