OX133, a monoclonal antibody recognizing protein-bound N-ethylmaleimide for the identification of reduced disulfide bonds in proteins

In vivo, enzymatic reduction of some protein disulfide bonds, allosteric disulfide bonds, provides an important level of structural and functional regulation. The free cysteine residues generated can be labeled by maleimide reagents, including biotin derivatives, allowing the reduced protein to be detected or purified. During the screening of monoclonal antibodies for those specific for the reduced forms of proteins, we isolated OX133, a unique antibody that recognizes polypeptide resident, N-ethylmaleimide (NEM)-modified cysteine residues in a sequence-independent manner. OX133 offers an alternative to biotin-maleimide reagents for labeling reduced/alkylated antigens and capturing reduced/alkylated proteins with the advantage that NEM-modified proteins are more easily detected in mass spectrometry, and may be more easily recovered than is the case following capture with biotin based reagents.

Emergence dynamics of barnyardgrass and jimsonweed from two depths when switching from conventional to reduced and no-till conditions

A cylinder experiment was conducted in northern Greece during 2005 and 2006 to assess emergence dynamics of barnyardgrass (Echinochloa crus-galli (L.) Beauv.) and jimsonweed (Datura stramonium L.) in the case of a switch from conventional to conservation tillage systems (CT). Emergence was surveyed from two burial depths (5 and 10 cm) and with simulation of reduced tillage (i.e. by soil disturbance) and no-till conditions. Barnyardgrass emergence was significantly affected by burial depth, having greater emergence from 5 cm depth (96%) although even 78% of seedlings emerged from 10 cm depth after the two years of study. Emergence of barnyardgrass was stable across years from the different depths and tillage regimes. Jimsonweed seeds showed lower germination than barnyardgrass during the study period, whereas its emergence was significantly affected by soil disturbance having 41% compared to 28% without disturbance. A burial depth x soil disturbance interaction was also determined, which showed higher emergence from 10 cm depth with soil disturbance. Jimsonweed was found to have significantly higher emergence from 10 cm depth with soil disturbance in Year 2. Seasonal emergence timing of barnyardgrass did not vary between the different burial depth and soil disturbance regimes, as it started in April and lasted until end of May in both years. Jimsonweed showed a bimodal pattern, with first emergence starting end of April until mid-May and the second ranging from mid-June to mid-August from 10 cm burial depth and from mid-July to mid-August from 5 cm depth, irrespective of soil disturbance in both cases.