While many variations of this procedure exist in the literature, in this chapter we describe the biotinylation protocol that we have applied for the investigation of quantitative changes in the cell surface expression and internalisation of ionotropic glutamate receptors in acute brain slices....
bLPL in addition also bound to low-density lipoprotein receptor related protein in ligand blotting. Thus, our protocol has produced biotinylated lipases which are both chemically and biologically active and can be used instead of iodinated lipases. 展开 ...
General protocols for the enzymatic incorporation of biotin–11–dUTP into DNA probes can be found inSection 1. A particularly interesting modification of the typical enzymatic incorporation protocol for biotin is described byDidenko (1993). The single-strand template is immobilized by adsorption onto ...
In principle, any scientist can perform this protocol within 3 days, although generating the proteomics data requires access to a high-end liquid chromatography–mass spectrometry setup. Data analysis and data visualization are relatively straightforward and can be performed using any type of software ...
(DNP) or a choice of several different fluorophores, as described below. Each of the protein labeling kits contains the preferred reactive dye or hapten—many of which have spacers to reduce interactions between the label and the biomolecule—along with a detaile...
This enabling technology not only eliminates the need for secondary detection reagents in many protocols, but also simplifies immunolabeling protocols that previously were time consuming or impractical, such as use of multiple antibodies derived from the same species...
The solid phase protocol for sulfhydryl labeling is significantly shorter than the corresponding solution phase method. Biotinylation in solid phase is convenient, efficient and easily applicable to small amounts of antibody (e.g. 100 microg). Antibody biotinylated on-column was found to be ...
Refers to protocol step 2b. All cells die, including conditions receiving virus. Potential solution You may not have succeeded in viral production, or the viral titer may not be high enough to sufficiently infect your cells. Consider re-making virus and measuring viral titer. Problem 3 Refers ...
Cells were first subjected to the ProtA-Turbo protocol described for fixed cells using the H3K9me3 antibody. After washing away the biotinylation reaction buffer, nuclei were resuspended in 1 ml of SDS buffer (50 mM Tris pH 8, 0.5% SDS, 100 mM NaCl, 5 mM EDTA) and incubated fo...
Protocol summary for the Cell Surface Protein Biotinylation and Isolation Kit Western blot analysis of enriched cell surface proteins Samples were prepared in duplicate from two cell lines using the Cell Surface Protein Biotinylation and Isolation Kit. Flow-through (F), elution (E), and bead boil ...