protoarray ® human protein microarrays v4.0  (Thermo Fisher)

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Interactome Mapping Uncovers a General Role for Numb in Protein Kinase Regulation *

doi: 10.1074/mcp.RA117.000114

Figure Lengend Snippet: A Numb PTB interactome identified by screening a protein microarray. A, B, Representative sections of a protein microarray (ProtoArray, Invitrogen) probed with the dNumb PTB domain. Protein spots that showed positive binding (red dots) were labeled. Control 1 spots represent Alexa Fluor 647 Rabbit anti-mouse IgG antibody; Control 2 spots represent Biotinylated anti-mouse antibody. C, A Venn diagram illustrating all potential binders of the dNumb or hNumb PTB domains identified from screening the ProtoArray and their overlap. D–F, Venn diagrams showing the protein kinases that bound the dNumb or/and the hNumb PTB domain.

Article Snippet: ProtoArray Hybridization The ProtoArray (Human protein microarray v4.0, Invitrogen, Cat. No. PAH0524011) was removed from −20 °C and placed immediately at 4 °C for at least 15 min.

Techniques: Microarray, Binding Assay, Labeling

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Interactome Mapping Uncovers a General Role for Numb in Protein Kinase Regulation *

doi: 10.1074/mcp.RA117.000114

Figure Lengend Snippet: Verification of Numb-PTB interactions by peptide array and in-solution binding. A, Binding profile for the dNumb PTB domain on an array of NxxF/Y motif-containing peptides. The peptides were derived from the PTB-interacting proteins identified using the ProtoArray (Fig. 2). B, Distribution of Z scores for the positive binders on the peptide array in (A). C–K, Binding curves of selected peptides from the peptide array for the dNumb PTB domain. See Table I for peptide sequences and Kd values.

Article Snippet: ProtoArray Hybridization The ProtoArray (Human protein microarray v4.0, Invitrogen, Cat. No. PAH0524011) was removed from −20 °C and placed immediately at 4 °C for at least 15 min.

Techniques: Peptide Microarray, Binding Assay, Derivative Assay

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Interactome Mapping Uncovers a General Role for Numb in Protein Kinase Regulation *

doi: 10.1074/mcp.RA117.000114

Figure Lengend Snippet: Binding affinities of peptides derived from several putative Numb-binding proteins identified in this study

Article Snippet: ProtoArray Hybridization The ProtoArray (Human protein microarray v4.0, Invitrogen, Cat. No. PAH0524011) was removed from −20 °C and placed immediately at 4 °C for at least 15 min.

Techniques: Binding Assay, Derivative Assay, Sequencing