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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
摘要

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).

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Sigma-Aldrich
脱氧核糖核酸酶 I 来源于牛胰腺, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein
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Lysozyme 来源于鸡蛋白, powder or granules, ≥39,000 units/mg protein
Sigma-Aldrich
过硫酸铵, for molecular biology, suitable for electrophoresis, ≥98%
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氢氧化钠, BioXtra, ≥98% (acidimetric), pellets (anhydrous)
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Triton X-100 溶液, BioUltra, for molecular biology, ~10% in H2O
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苯甲磺酰氟, ≥98.5% (GC)
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酵母提取物, for use in microbial growth medium
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碘乙酰胺, ≥99% (NMR), crystalline
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牛血清白蛋白 来源于牛血清, lyophilized powder, crystallized, ≥98.0% (GE)
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叠氮化钠, BioUltra, ≥99.5% (T)
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D-乳糖 一水合物, ≥99% (HPLC), BioUltra
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Trizma® 盐酸盐 盐酸盐, BioUltra, for molecular biology, ≥99.0% (AT)
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十二烷基硫酸钠, ACS reagent, ≥99.0%
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氯化钠, BioXtra, ≥99.5% (AT)
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IPTG, ≥99% (TLC), ≤0.1% Dioxane
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α-乳糖-琼脂糖, saline suspension
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大豆蛋白胨, 酶消化物 来源于酪蛋白, Pancreatic digest of casein