Luminescent detection method for immunodot, Western, and Southern blots.

Abstract

An anti-peroxidase-anti-biotin hybrid hybridoma rat cell line, capable of producing a bispecific monoclonal antibody, has been derived to explore its use in conjunction with a luminol immunodetection system. Luminescence was detected using x-ray film. The method was sufficiently sensitive and effective, but was less sensitive than autoradiographic methods using high-specific-activity 32P-labeled probes. Exposure times, on the other hand, were of the order of seconds rather than days. The direct binding of both peroxidase and biotin by the bispecific monoclonal antibody is simpler but less sensitive than the more conventional indirect method using a commercial peroxidase coupled with anti-rat antibody as a developing antibody.

Complications in cell-surface labelling by biotinylation of Candida albicans due to avidin conjugate binding to cell-wall proteins

“…The membranes were washed three times (10 min) in 50 mM Tris, pH 7n5. Avidin or antibody binding was detected by 3,3h-diaminobenzidinejH # O # or by enhanced chemiluminescence (Leong et al, 1986). Blots were scanned on a flat-bed scanner and the image quality (contrast and brightness) was adjusted using Photoshop (version 6.0 ; Adobe) for reproduction.…”

New Approaches to the Detection of Microbial Plant Pathogens

“…An adaptation of this approach, using different reagents, is the chemiluminescence approach (11) for the detection of a nucleic acid sequence (Thorpe andKricka. 1987: Urdea et ai., 1988 hybridizations (Baldwin, Holzman and Holzman, 1986;Leong, Milstein and Pannell, 1986;Tanaka and Ishikawa, 1986;Terouanne et al, 1986Terouanne et al, , 1987Brochu etal., 1987;Geiger and Miska, 1987;Hauber and Geiger, 1988;Kricka, 1988) with great sensitivity ( Figure 5A). The signal can be measured with a liquid scintillation counter (Nguyen, Morange and Bensaude, 1988) or recorded on photographic film.…”

In Staphylococcus aureus, the Particulate State of the Cell Envelope Is Required for the Efficient Induction of Host Defense Responses

“…Immunodot blotting. To confirm the presence of LTA in the SCEs or the detection of calprotectin in peritoneal cell lysates (PCLs) and peritoneal fluids (PFs), the immunodot blotting was carried out as described previously (51). Briefly, after activation of a PVDF membrane with methanol, samples were mixed with ethanol (final 50% concentration), spotted on the membrane, and dried.…”