Immunoenzymatic method for detection of terminal deoxynucleotidyl transferase by light and electron microscopy.
Methods for using rabbit polyclonal and mouse monoclonal antibodies to detect terminal deoxynucleotidyl transferase (TdT) in human leukemia lines were developed. Use of 1% paraformaldehyde in 0.1 M Na cacodylate buffer, pH 7.6, as a fixative provides a procedure that can be used for immunofluorescence, immunoalkaline phosphatase (APAAP), or horseradish peroxidase conjugate detection procedures in these cell lines. The peroxidase procedure can also be used for electron microscopy, clearly demonstrating nuclear localization of TdT in interphase nuclei of TdT-positive cells.
Di Primio
R
R
Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-5044.
Bollum
F J
FJ
D000469:Alkaline Phosphatase; D000818:Animals; D000911:Antibodies, Monoclonal; D004253:DNA Nucleotidylexotransferase / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006735:Horseradish Peroxidase; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G; D051379:Mice; D008854:Microscopy, Electron; D011817:Rabbits; D014407:Tumor Cells, Cultured / Q000201:enzymology* / Q000473:pathology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; C051538:IGG-horseradish peroxidase; D007074:Immunoglobulin G; D006735:Horseradish Peroxidase; D004253:DNA Nucleotidylexotransferase; D000469:Alkaline Phosphatase