SDS-PAGE Analysis of Recombinant Proteins

The purified recombinant proteins (LcBSH and GfBSH) were treated with 4× premixed sample buffer solution (Bio-Rad, Hercules, CA, USA) and heat-denatured at 95 °C for 5 min using a thermal cycler (TaKaRa). The resulting protein solutions were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using Mini-PROTEAN TGX precast polyacrylamide gels (Bio-Rad), according to the methods described in previous studies [23 (link),24 (link),25 (link),54 (link)]. The gels were stained overnight with QC Colloidal Coomassie Stain (Bio-Rad), with gentle agitation, then decolorized with deionized water.

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Morinaga K., Kusada H, & Tamaki H. (2022). Bile Salt Hydrolases with Extended Substrate Specificity Confer a High Level of Resistance to Bile Toxicity on Atopobiaceae Bacteria. International Journal of Molecular Sciences, 23(18), 10980.

Publication 2022

thermal cycler Mini-PROTEAN TGX precast polyacrylamide gels QC Colloidal Coomassie Stain

Buffer Gels Polyacrylamide gels Protein Recombinant proteins Sds page Stain

Corresponding Organization : University of Tsukuba

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Variable analysis

independent variables

Treatment of purified recombinant proteins (LcBSH and GfBSH) with 4× premixed sample buffer solution
Heat denaturation of protein solutions at 95 °C for 5 min

dependent variables

Characteristics of the analyzed protein solutions (e.g., molecular weight, purity)

control variables

Use of Mini-PROTEAN TGX precast polyacrylamide gels (Bio-Rad) for SDS-PAGE
Staining of the gels overnight with QC Colloidal Coomassie Stain (Bio-Rad) and decolorizing with deionized water

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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