- PA5-82152 - Invitrogen Antibodies TRIM33 antibody

Mojsa B, Tatham MH, Davidson L, Liczmanska M, Branigan E, Hay RT
Molecular & cellular proteomics : MCP 2021;20:100164

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of TRIM33 by a TRIM33 polyclonal antibody (Product # PA5-82152). Analysis in mouse cell line NIH-3T3 and rat cell line NBT-II.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: CRISPR-Cas9 mediated genome editing ofTRIM33 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR1029102_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of TRIM33 was performed by loading 30 µg of MCF7 Cas9 (Lane 1) and MCF7 Cas9 cells transduced with TRIM33 Lentiviral sgRNA (Lane 2) modified whole cell extracts. The samples were electrophoresed using NuPAGE™ 3 to 8%, Tris-Acetate, 1.0 mm, Mini Protein Gel (Product # EA03755BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with TRIM33 Polyclonal Antibody (Product # PA5-82152) using 1:2,500 dilution and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036 1:4,000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toTRIM33 (Fig (b)).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of TRIM33 by a TRIM33 polyclonal antibody (Product # PA5-82152). Analysis in human cell lines MCF-7 and HeLa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of TRIM33 in U-251 MG cells using a TRIM33 polyclonal antibody (Product # PA5-82152). The analysis shows localization to nucleoplasm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of TRIM33 in human hippocampus using a TRIM33 polyclonal antibody (Product # PA5-82152). The analysis shows strong nuclear positivity in neuronal cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 Functions of SUMO1 and SUMO2 targets in hiPSCs. A , in total, 976 SUMO sites were identified from 6His-SUMO1-KGG and 6His-SUMO2-KGG IPS cells, of which 155 were novel compared with previous high-throughput SUMO site proteomics studies ( supplemental File S2 ). Proteins with three or more novel sites are highlighted. B , summary of the top 50 SUMO substrates by total SUMO1+SUMO2 GGK-peptide intensity for all identified sites. Gene names are shown with numbers of sites in brackets . Bars are color-coded by category shown in panel D . Insert shows contribution to total GGK peptide intensity of proteins from the categories shown in ( D ) (note categories are not mutually exclusive). C , immunoblot analysis of ChIPS4 cells treated with the SUMO E1 conjugating enzyme inhibitor ML792 or DMSO for 48 h. Total protein extracts were probed with anti-TRIM28, anti-TRIM24, anti-CTCF, anti-DNMT3b, anti-SALL4, anti-TRIM33, and anti-tubulin or anti-Lamin A/C antibodies (loading controls). SUMO-modified species above the band for unmodified proteins (*) reduce with ML792 treatment. D , STRING interaction network of the 427 IPS SUMO substrates. Only high confidence interactions were considered from ""Text mining,"" ""Experiments,"" and ""Databases"" sources. Network PPI enrichment p -value

PA5-82152