rabbit polyclonal anti-usp7  (Novus Biologicals)

Journal: eLife

Article Title: The termination of UHRF1-dependent PAF15 ubiquitin signaling is regulated by USP7 and ATAD5

doi: 10.7554/eLife.79013

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-USP7 (Rabbit polyclonal) , Thermo Fisher Scientific , Cat# A300-033A, RRID: AB_203276 , WB (1:1000).

Techniques: Recombinant, Expressing, Purification, Bacteria, Ubiquitin Proteomics, Methylation

Journal: International Journal of Molecular Sciences

Article Title: A Proteomic Approach for Systematic Mapping of Substrates of Human Deubiquitinating Enzymes

doi: 10.3390/ijms22094851

Figure Lengend Snippet: Isolation of ubiquitinated material by biotin pull-downs in siRNA silenced samples. ( A ) Analysed deubiquitinating enzymes (DUBs). The reduction levels achieved for each of the DUB analysed are shown. USP1, USP7, USP9X and USP11 levels were detected by Western blot, whereas RT-qPCR was used for USP42. The mRNA levels were normalized to the control samples. The average mRNA level in USP42-silenced sample (0.65) is shown with a dashed line. ( B ) Silver staining of the material purified by biotin-pulldowns. Just a few endogenous carboxylases proteins are detected in samples overexpressing only the BirA enzyme (BirA sample). However, a smear corresponding to ubiquitinated material is detected when the BirA enzyme and the biotinylated ubiquitins are co-expressed together ( bio Ub samples). Similar amount of ubiquitinated material is isolated from cell treated with the siRNA control (−) and a DUB-specific siRNA (+). Avidin monomers and dimers are indicated with arrowheads. The four slices that are extracted from each sample for MS analysis are highlighted with the brackets. Only the bio Ub samples were analysed using the LFQ MS approach.

Article Snippet: The following primary antibodies were used: goat anti biotin-horseradish peroxidase (HRP)-conjugated antibody (Cell Signaling Technology, Danvers, MA, USA; catalogue number 7075) at 1:1000; USP1 (Cell Signaling Technology, Danvers, MA, USA; catalogue number 8033); rabbit polyclonal anti-USP7 antibody (Bethyl laboratories, Montgomery, TX, USA; catalogue number A300-033A) at 1:3000; rabbit polyclonal anti-USP9X antibody (ProteinTech, Rosemont, IL, USA; catalogue number 55054-1-AP) at 1:1000; rabbit monoclonal anti-USP11 (Abcam, Cambridge, UK; catalogue number ab109232); mouse monoclonal anti-Tubulin beta antibody (Developmental Studies Hybridoma Bank, Iowa City, IA, USA; catalogue number E7) at 1:1000).

Techniques: Isolation, Western Blot, Quantitative RT-PCR, Control, Silver Staining, Purification, Avidin-Biotin Assay

Journal: International Journal of Molecular Sciences

Article Title: A Proteomic Approach for Systematic Mapping of Substrates of Human Deubiquitinating Enzymes

doi: 10.3390/ijms22094851

Figure Lengend Snippet: Putative deubiquitinating enzyme (DUB) substrates.

Article Snippet: The following primary antibodies were used: goat anti biotin-horseradish peroxidase (HRP)-conjugated antibody (Cell Signaling Technology, Danvers, MA, USA; catalogue number 7075) at 1:1000; USP1 (Cell Signaling Technology, Danvers, MA, USA; catalogue number 8033); rabbit polyclonal anti-USP7 antibody (Bethyl laboratories, Montgomery, TX, USA; catalogue number A300-033A) at 1:3000; rabbit polyclonal anti-USP9X antibody (ProteinTech, Rosemont, IL, USA; catalogue number 55054-1-AP) at 1:1000; rabbit monoclonal anti-USP11 (Abcam, Cambridge, UK; catalogue number ab109232); mouse monoclonal anti-Tubulin beta antibody (Developmental Studies Hybridoma Bank, Iowa City, IA, USA; catalogue number E7) at 1:1000).

Techniques: Ubiquitin Proteomics