{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,3]],"date-time":"2025-11-03T08:59:21Z","timestamp":1762160361438},"reference-count":23,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2005,2,4]],"date-time":"2005-02-04T00:00:00Z","timestamp":1107475200000},"content-version":"vor","delay-in-days":5148,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cell Motil. Cytoskeleton"],"published-print":{"date-parts":[[1991,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Monoclonal antibodies were raised against calmodulin purified from <jats:italic>Dictyostelium discoideum.<\/jats:italic> To increase its antigenicity, the calmodulin was conjugated to keyhole limpet hemocyanin; mice were immunized with the conjugate. Hybridomas producing antibodies against calmodulin were identified by screening culture supernatants with calmodulin coupled to bovine serum albumin. The specificity of antibodies from hybridoma culture supernatants was tested by Western blot of <jats:italic>Dictyostelium<\/jats:italic> cell lysates. For this purpose, methods were developed that permitted sensitive detection of calmodulin bound to membranes. The key elements of the blotting protocol were use of PVDF membrane, transfer conducted in phosphate buffer, and glutaraldehyde fixation after transfer. These methods permitted detection of as little as 0.1 ng of calmodulin spotted directly onto the membrane, or 10 ng transferred from an SDS polyacrylamide gel. Ten calmodulin\u2010specific antibodies were identified; most of these reacted preferentially with the calcium\u2010containing form of <jats:italic>Dictyostelium<\/jats:italic> calmodulin. Several of the monoclonal antibodies cross\u2010reacted with calmodulin from bovine brain.<\/jats:p>","DOI":"10.1002\/cm.970180206","type":"journal-article","created":{"date-parts":[[2005,2,24]],"date-time":"2005-02-24T03:04:46Z","timestamp":1109214286000},"page":"113-122","source":"Crossref","is-referenced-by-count":75,"title":["Production and specificity of monoclonal antibodies against calmodulin from dictyostelium discoideum"],"prefix":"10.1002","volume":"18","author":[{"given":"Durvis","family":"Hulen","sequence":"first","affiliation":[]},{"given":"Andre","family":"Baron","sequence":"additional","affiliation":[]},{"given":"Jeffrey","family":"Salisbury","sequence":"additional","affiliation":[]},{"given":"Margaret","family":"Clarke","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2005,2,4]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/cm.970020506"},{"key":"e_1_2_1_3_1","first-page":"377","volume-title":"Calcium as a Second Messenger in Eukaryotic Microbes","author":"Clarke M.","year":"1990"},{"key":"e_1_2_1_4_1","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1128\/jb.141.1.397-400.1980","article-title":"Isolation and properties of calmodulin from Dictyostelium discoideum","volume":"141","author":"Clarke M.","year":"1980","journal-title":"J. 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