In biomedical research, indirect immunofluorescence labelling by use of primary and secondary antibodies is central for revealing the spatial distribution of multiple cellular antigens. However, labelling is regularly restricted to few antigens since species variation of primary and corresponding secondary an...
MCF-7 cells were infected with CRISPR sgRNA targeting individual VGLL3 binding partners or in combination with sgRNA targeting LATS1/2. m VGLL3 interacts with NCOR2. MCF-7 cells expressing Flag-VGLL3, or control vector, were immunoprecipitated with Flag antibody. Western blotting for co-...
When the fluorescently labeled peptide or antibody is exposed to light of the proper wave length, its presence can then be detected due to fluorescence. Among the most commonly used fluorescent labelling compounds are fluorescein isothiocyanate, rhodamine, phycoerythrin, phycocyanin, allophycocyanin, o-...
Samples were quenched with 50 mM NH4Cl, blocked in PBS with 1% BSA and immunolabeled with a-hPRXD6 1A11 (SantaCruz, 1:20) IgG1a monoclonal antibody in PBS with 1% BSA followed by a bridge step labelling with a polyclonal Rabbit Anti...
This invention relates to methods for the evaluation and/or quantification of the binding affinity of small molecules or other compounds to target components contained within an analyte, such as target proteins contained within the proteome of a cell or tissue.Inventors...
Most current computational methods used to design proteins that bind to a target surface utilize information derived from structures of the native complex on specific side-chain interactions or protein backbone placements optimal for binding1,2,3. Computational docking of antibody scaffolds with varied ...
and can thus be captured, resulting in a low yield.7In addition, recovering fully intact exosomes can be difficult after antibody binding in immunoaffinity isolation.82 Zarovniet al.evaluated several commercially available kits for immunoaffinity-based isolation and modified their protocols to increase ...
purification; BSA as a control, and fractions of BslA and TasA that were obtained during purification. Middle: immunoblot of purified proteins using an anti-TasA antibody (1:20,000). Right: far-western blot using an anti-TasA antibody (1:20,000) after renaturing of the proteins and incuba...
A variant of these methods is DNA-PAINT12, which builds on repetitive and reversible binding of fluorophore-labeled imager-strands of single stranded DNA to a complementary target strand, which in case of protein labeling is tagged to a protein-specific antibody, nanobody or small molecule such ...
FIG. 2. Binding affinity of the REST phosphomimetic peptide (RPP) for CTDSP1 was measured using the Monolith (NanoTemper). The affinity of RPP (Table 4; SEQ ID NO: 165) for CTDSP1 (Table 2; SEQ ID NO:163) is approximately 130 pM. Binding was assessed by fluorescently labelling the Hi...