chromotek histone label atto488  (Proteintech)

Journal: Science Advances

Article Title: Insights into the structure and evolution of the human SAGA complex by affinity-ligand purification

doi: 10.1126/sciadv.aec8104

Figure Lengend Snippet: ( A ) Proteomic analysis of the purified SAGA and ATAC complexes. For each identified protein the table shows the normalized spectral abundance factor (NSAF) value calculated from the PSMs divided by the number of amino acids (AA), and the normalization of the NSAF value to SUPT20H as rough estimation of stoichiometry. SAGA subunits are colored purple, ATAC subunits are colored blue, purple bold represents the common subunits between SAGA and ATAC, and copurified bromodomain containing proteins are in green. ( B ) Western blot analysis of different purification steps detecting the TADA2B subunit of SAGA. ( C ) Relative quantification of the detected bands in (B). ( D ) Colloidal Coomassie blue–stained SDS-PAGE of the purified SAGA and ATAC complexes; the identity of indicated bands was verified by MS. ( E ) Deubiquitination activity of the purified SAGA on the fluorogenic ubiquitin–amino methyl coumarin (Ub-AMC) over time. ( F ) Acetyltransferase activity of the purified SAGA and ATAC complex on nucleosome core particles (NCPs). Acetylation of histone was detected by a pan-acetyl-lysine antibody. Anti-GCN5 and histone H2B antibodies were used as loading control of SAGA/ATAC and NCPs, respectively.

Article Snippet: The primary antibodies used are against TADA2B (produced in-house, no. 3122), GCN5 (produced in-house, no. 2676), and histone H2B (Cell Signaling, 2934S).

Techniques: Purification, Western Blot, Quantitative Proteomics, Staining, SDS Page, Activity Assay, Ubiquitin Proteomics, Control