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addgene plasmids 63571  (Addgene inc)


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    Addgene inc addgene plasmids 63571
    Addgene Plasmids 63571, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/addgene plasmids 63571/product/Addgene inc
    Average 93 stars, based on 13 article reviews
    addgene plasmids 63571 - by Bioz Stars, 2026-03
    93/100 stars

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    Purification of Ubiquitin and UBA1 <t>(E1)</t> (A) The samples of GST-Ubiquitin affixed to GST resin, of flowthrough (FT) after PreScission protease digestion and of elution fractions from size exclusion chromatography (SEC) (3 μL) were analyzed by SDS-PAGE and Coomassie staining. (B) Bound UBA1 protein (3 μL) and elution fractions (3 μL) from GST-Ub affinity resin were analyzed by SDS-PAGE and Coomassie staining. (C) The UBA1 (E1) elution fractions from size exclusion chromatography (SEC) (3 μL) were analyzed by SDS-PAGE and Coomassie staining.
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    Purification of Ubiquitin and UBA1 (E1) (A) The samples of GST-Ubiquitin affixed to GST resin, of flowthrough (FT) after PreScission protease digestion and of elution fractions from size exclusion chromatography (SEC) (3 μL) were analyzed by SDS-PAGE and Coomassie staining. (B) Bound UBA1 protein (3 μL) and elution fractions (3 μL) from GST-Ub affinity resin were analyzed by SDS-PAGE and Coomassie staining. (C) The UBA1 (E1) elution fractions from size exclusion chromatography (SEC) (3 μL) were analyzed by SDS-PAGE and Coomassie staining.

    Journal: STAR Protocols

    Article Title: Protocol for evaluating the E3 ligase activity of BRCA1-BARD1 and its variants by nucleosomal histone ubiquitylation

    doi: 10.1016/j.xpro.2024.103294

    Figure Lengend Snippet: Purification of Ubiquitin and UBA1 (E1) (A) The samples of GST-Ubiquitin affixed to GST resin, of flowthrough (FT) after PreScission protease digestion and of elution fractions from size exclusion chromatography (SEC) (3 μL) were analyzed by SDS-PAGE and Coomassie staining. (B) Bound UBA1 protein (3 μL) and elution fractions (3 μL) from GST-Ub affinity resin were analyzed by SDS-PAGE and Coomassie staining. (C) The UBA1 (E1) elution fractions from size exclusion chromatography (SEC) (3 μL) were analyzed by SDS-PAGE and Coomassie staining.

    Article Snippet: The plasmids of pFastbac-Flag-BRCA1 (modified based on the plasmid from Jeffrey Parvin; deposited in Addgene (#223228)) and pFastBac-TwinStrepTag-BARD1 (Addgene#137166) are used for Flag-BRCA1 and Twin-StrepTag-BARD1 expression in insect cells., c. The expression plasmids for human E1 (pET3a-hUBA1 (Addgene#63571)), E2s (pET28a-UBE2D3 (Addgene#12643), pET15-UBE2W, pET24a-UBE2E3, pET24a-UBE2E2, pET24a-UBE2E1, pET24a-UBE2N, pET24a-UBE2V2), and Ubiquitin (pET15-Ub (Addgene#12647)) were described before., , , Plasmids of pDEST17-UBE2B (Addgene#15781) was from Addgene.

    Techniques: Purification, Ubiquitin Proteomics, Size-exclusion Chromatography, SDS Page, Staining

    Reagents for the in vitro ubiquitination assay

    Journal: STAR Protocols

    Article Title: Protocol for evaluating the E3 ligase activity of BRCA1-BARD1 and its variants by nucleosomal histone ubiquitylation

    doi: 10.1016/j.xpro.2024.103294

    Figure Lengend Snippet: Reagents for the in vitro ubiquitination assay

    Article Snippet: The plasmids of pFastbac-Flag-BRCA1 (modified based on the plasmid from Jeffrey Parvin; deposited in Addgene (#223228)) and pFastBac-TwinStrepTag-BARD1 (Addgene#137166) are used for Flag-BRCA1 and Twin-StrepTag-BARD1 expression in insect cells., c. The expression plasmids for human E1 (pET3a-hUBA1 (Addgene#63571)), E2s (pET28a-UBE2D3 (Addgene#12643), pET15-UBE2W, pET24a-UBE2E3, pET24a-UBE2E2, pET24a-UBE2E1, pET24a-UBE2N, pET24a-UBE2V2), and Ubiquitin (pET15-Ub (Addgene#12647)) were described before., , , Plasmids of pDEST17-UBE2B (Addgene#15781) was from Addgene.

    Techniques: In Vitro, Ubiquitin Proteomics, Concentration Assay