alphascreen flag m2 detection kit  (Revvity)

Journal: iScience

Article Title: A series of xanthenes inhibiting Rad6 function and Rad6-Rad18 interaction in the PCNA ubiquitination cascade

doi: 10.1016/j.isci.2022.104053

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Article Snippet: AlphaScreen FLAG (M2) detection kit , PerkinElmer , Cat#6760613.

Techniques: Virus, Recombinant, Ubiquitin Proteomics, Modification, Amplified Luminescent Proximity Homogenous Assay, Plasmid Preparation, Software

Journal: Assay and Drug Development Technologies

Article Title: Development of a High-Throughput Screening-Compatible Assay for the Discovery of Inhibitors of the AF4-AF9 Interaction Using AlphaScreen Technology

doi: 10.1089/adt.2012.495

Figure Lengend Snippet: Table 5.

Article Snippet: Gray, 384-well AlphaPlates SW, 7.5% bovine serum albumin (BSA), and AlphaScreen FLAG detection kit (Catalog no. 6760613C) containing anti-FLAG-coated acceptor beads and streptavidin donor beads were purchased from Perkin Elmer.

Techniques: Amplified Luminescent Proximity Homogenous Assay

Journal: Journal of Biological Chemistry

Article Title: Identification of Binding Sites for Myeloid Differentiation Primary Response Gene 88 (MyD88) and Toll-like Receptor 4 in MyD88 Adapter-like (Mal)

doi: 10.1074/jbc.m112.415810

Figure Lengend Snippet: FIGURE 6. Mapping of the effect of 38 selected Mal prey mutations on Mal-MalinteractioninAlphaScreenassays.Residuesthatcorrespondto38 tested mutants are colored according to the effect on the AlphaScreen assay, as indicated in the color scale in panel A. Other residues are colored black. Backboneatomsarecoloredgray.ADTTmoleculelinkedtoCys-157iscolored blue. A, shown is mapping of the mutants on a Mal monomer, oriented as in Fig. 2A. Several mutations that affect Mal-Mal interaction are found in the dimer interface. B, mapping of the mutants on a Mal dimer is shown. The dimer interface is indicated by a dashed line. Residues that affect the Mal-Mal interactionoutsidethesymmetricalinterface(red/orange)areeitherburiedor are isolated and surrounded by mutations that do not affect the Mal-Mal AlphaScreen assay.

Article Snippet: The lysates were incubated for 2 h at room temperature with biotinylated anti-E-tag antibody (Amersham Biosciences), subsequently for 1 h with the AlphaScreen FLAGTM (M2) detection kit (PerkinElmer Life Sciences) acceptor beads, and finally for 30 min with streptavidin donor beads.

Techniques: Amplified Luminescent Proximity Homogenous Assay, Isolation

Journal: Journal of Biological Chemistry

Article Title: Identification of Interaction Sites for Dimerization and Adapter Recruitment in Toll/Interleukin-1 Receptor (TIR) Domain of Toll-like Receptor 4

doi: 10.1074/jbc.m111.282350

Figure Lengend Snippet: FIGURE 5. Top, AlphaScreenTM analysis of the interaction between TLR4ic mutants and TRAM. HEK293T cells were transiently co-transfected with plas- mids encoding the E-tagged TLR4 WT or mutants and FLAG-tagged TRAM. The transfected cells were lysed after 48 h, and protein interactions were detected with the AlphaScreen FLAGTM (M2) detection kit (PerkinElmer Life Sciences). The results are displayed as fold induction, namely relative to the counts of an irrelevant E-tagged construct. Bottom, expression levels of the E-tagged TLR4 constructs were detected on Western blot; TLR4 is the lower band on the blot. In the 1st lane (negative control (neg. ctrl)), the control lysate was loaded; the order of the other loaded samples aligns with the order of the samples on the graph.

Article Snippet: The lysates were incubated for 2 h at room temperature with biotinylated anti-E-tag antibody (Amersham Biosciences) and subsequently for 2 h with the AlphaScreenTM FLAGTM (M2) detection kit (PerkinElmer Life Sciences) acceptor beads.

Techniques: Transfection, Amplified Luminescent Proximity Homogenous Assay, Construct, Expressing, Western Blot, Negative Control, Control