kinase domains ephb1  (Addgene inc)

Journal: eLife

Article Title: High-throughput profiling of sequence recognition by tyrosine kinases and SH2 domains using bacterial peptide display

doi: 10.7554/eLife.82345

Figure Lengend Snippet: ( A ) Heatmaps depicting the specificities of c-Abl, Fer, EPHB1, and EPHB2. Enrichment scores were log 2 -transformed and are displayed on a color scale from blue (disfavored sequence features, negative value), to white (neutral sequence features, near zero value), to red (favored sequence features, positive value). Values in the heatmaps are the average of three replicates. ( B ) Sequences of consensus peptides identified through X 5 -Y-X 5 screens, compared with previously reported SrcTide and AblTide sequences. ( C ) Phosphorylation kinetics of five consensus peptides against five kinases. Initial rates were normalized to the rate of the cognate consensus peptide. All peptides were used at a concentration of 100 μM, and the kinases were used at a concentration of 10–50 nM. Error bars represent the standard deviation from at least three measurements. Figure 2—source data 1. Position-specific amino acid enrichment matrices from the tyrosine kinase X 5 -Y-X 5 library screens. Matrices calculated with and without inclusion of multi-tyrosine sequences are provided.

Article Snippet: Recombinant DNA reagent , pET-His6-TEV-EPHB1(KD) , PMID: 30004690 , Addgene: 79694 , bacterial expression vector encoding the human EPHB1 kinase domain (residues 602–896) with an N-terminal His6-tag and TEV protease recognition sequence.

Techniques: Transformation Assay, Sequencing, Concentration Assay, Standard Deviation

Journal: eLife

Article Title: High-throughput profiling of sequence recognition by tyrosine kinases and SH2 domains using bacterial peptide display

doi: 10.7554/eLife.82345

Figure Lengend Snippet: Cells displaying the X 5 -Y-X 5 library were treated with a kinase cocktail containing c-Src, c-Abl, AncSZ, and EPHB1 for 3 hr, then labeled with a pan-phosphotyrosine antibody (PY20 PerCP-eFluor 710) and analyzed by flow cytometry.

Article Snippet: Recombinant DNA reagent , pET-His6-TEV-EPHB1(KD) , PMID: 30004690 , Addgene: 79694 , bacterial expression vector encoding the human EPHB1 kinase domain (residues 602–896) with an N-terminal His6-tag and TEV protease recognition sequence.

Techniques: Labeling, Flow Cytometry

Journal: eLife

Article Title: High-throughput profiling of sequence recognition by tyrosine kinases and SH2 domains using bacterial peptide display

doi: 10.7554/eLife.82345

Figure Lengend Snippet:

Article Snippet: Recombinant DNA reagent , pET-His6-TEV-EPHB1(KD) , PMID: 30004690 , Addgene: 79694 , bacterial expression vector encoding the human EPHB1 kinase domain (residues 602–896) with an N-terminal His6-tag and TEV protease recognition sequence.

Techniques: Clone Assay, Over Expression, Strep-tag, Recombinant, Plasmid Preparation, Sequencing, Variant Assay, Expressing, Mutagenesis, Marker, Amplification, Synthesized, Western Blot, Purification, Software