used to affinity purify all goat antibodies that bind to mouse IgG1. These anti-mouse IgG1 antibodies would then be further purified by passage through a chromatography column(s) containing mouse IgG2a, IgG2b, IgG3, IgM, etc., to remove any an...
Our approach relies upon the expression and purification of the non-phosphorylated phosphosite libraries and an active kinase of interest. The kinase is then reacted with the phosphosite library in vitro, enabling a proteome-wide phosphosite substrate screen (Fig. 4a). Reactions are subsequently ...
Briefly, 0.1 µM of enzymes were allowed to react with 1 µM me7GpppA-capped 25 nt RNA in a buffer (50 mM Tris-HCl pH 8.0, 5 mM KCl, 1 mM dithiothreitol, 0.2 mM SAM) supplemented with 1 mM MgCl2 or MnCl2, or CaCl2 or 5 mM EDTA. The reactions ...
The mechanism of some enzymes makes it difficult to obtain a continuous optical change during reaction with an enzyme substrate. However, a discontinuous assay can often be developed by derivatizing the reaction products with one of the reagents described in Fl...
To prevent increased HIF activity in normoxia, HIFs are tightly regulated by different pathways and enzymes. HIFs undergo proline hydroxylation, ubiquitination, SUMOylation, S-nitrosylation, asparagine hydroxylation and phosphorylation to promote HIF degradation. One of the major HIF regulatory proteins ...
Besides, our previous study about proteins and their quinones adducts showed that many natural proteins and enzymes, that could be candidate targeted analytes for immunoassay, such as cytochrome c, alcohol dehydrogenase,β-lactoglobulin, lysozyme, and HSA, does not show any CL response to the DTT ...
(ET) signaling pathway is then activated, which can quickly initiate the plant defense response that acts in a positive manner in response to pathogens, such as through the activation of detoxification enzymes, antimicrobial proteins, pathogenesis-related protein (PR), cell wall reinforcement [16, ...
for example quenching of singlet oxygen and non-photochemical quenching of absorbed energy (Komenda and Sobotka, 2016); the second one are nonenzimatic antioxidants like α-tocopherol or carotenoids; and the last one to control the level of ROS are the antioxidant enzymes (Latifi et al., 2009...
Replicative DNA polymerases cannot initiate DNA synthesis de novo and rely on dedicated RNA polymerases, primases, to generate a short primer. This primer is then extended by the DNA polymerase. In diverse archaeal species, the primase has long been know
enzymes can promote lipid peroxidation and ferroptosis. Future investigations are needed to determine the potential involvement of other oxygenases, such as cyclooxygenases and peroxygenases, in lipid peroxidation. Non-enzymatic lipid peroxidation of PUFA-PLs is driven by the Fenton reaction, with iron...