Linked Life Data

12505721

Source:http://linkedlifedata.com/resource/pubmed/id/12505721

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-2
pubmed:dateCreated
2002-12-30
pubmed:abstractText
By employing two different immunogens and two different antibody-screening strategies, we established four mouse hybridoma clones producing monoclonal antibodies against N epsilon -acetyllysine. Three different protocols were used in this study; i.e., mice were (1) immunized with an N epsilon -acetyllysine-containing peptide, Gly-Lys(Ac)- epsilon -aminocaproic acid (Aca)-Cys, conjugated to KLH, and the hybridoma clones were screened for their reactivity to a histone H3 peptide containing five acetyllysines; (2) immunized as in "1" and screened with chemically acetylated bovine serum albumin (BSA); (3) immunized with chemically acetylated keyhole limpet hemocyanin (KLH) and screened with chemically acetylated BSA. Antibodies produced by the four different hybridomas established here all reacted with acetyllysine residues, but their reactivity was not the same when evaluated with enzyme-linked immunosorbent assay (ELISA), Western blotting, and resonant mirror sensor analyses. Among the three protocols examined, protocol "3" was especially useful to obtain hybridomas producing anti-N epsilon -acetyllysine antibodies that could detect not only the acetylated histones but also other acetylated proteins. By cloning and sequencing the cDNAs encoding the variable regions of the antibodies, we found that their framework sequences were almost the same, which suggests that some framework amino acids in addition to their complementarity determining regions (CDRs) directly contribute to their recognition function.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0022-1759
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
272
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
161-75
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:12505721-Acetylation, pubmed-meshheading:12505721-Amino Acid Sequence, pubmed-meshheading:12505721-Animals, pubmed-meshheading:12505721-Antibodies, Monoclonal, pubmed-meshheading:12505721-Antibody Specificity, pubmed-meshheading:12505721-Antigen-Antibody Reactions, pubmed-meshheading:12505721-Antigens, pubmed-meshheading:12505721-Base Sequence, pubmed-meshheading:12505721-Blotting, Western, pubmed-meshheading:12505721-Cloning, Molecular, pubmed-meshheading:12505721-Complementarity Determining Regions, pubmed-meshheading:12505721-DNA, Complementary, pubmed-meshheading:12505721-Electrophoresis, Gel, Two-Dimensional, pubmed-meshheading:12505721-Enzyme-Linked Immunosorbent Assay, pubmed-meshheading:12505721-Hybridomas, pubmed-meshheading:12505721-Immunoglobulin Variable Region, pubmed-meshheading:12505721-Kinetics, pubmed-meshheading:12505721-Lysine, pubmed-meshheading:12505721-Mice, pubmed-meshheading:12505721-Molecular Sequence Data, pubmed-meshheading:12505721-Molecular Structure, pubmed-meshheading:12505721-Precipitin Tests, pubmed-meshheading:12505721-Proteins, pubmed-meshheading:12505721-Sequence Homology, Nucleic Acid
pubmed:year
2003
pubmed:articleTitle
Four different clones of mouse anti-acetyllysine monoclonal antibodies having different recognition properties share a common immunoglobulin framework structure.
pubmed:affiliation
CREST Research Project, Japan Science and Technology Corporation, Kawaguchi Center Building, 4-1-8 Hon-cho, Saitama 332-0012, Japan. gc5y-kmt@asahi-net.or.jp
pubmed:publicationType
Journal Article, Comparative Study, In Vitro