Procathepsin L-specific antibodies that recognize procathepsin L but not cathepsin L.
Procathepsin L was purified to apparent homogeneity from the culture medium of v-Ha-ras transformed NIH3T3 (Ras-NIH) cells in three steps; anion-exchange chromatography, gel filtration, and re-gel filtration. SDS-PAGE analyses revealed that the purified samples contained only the precursor form, procathepsin L, but not the mature enzyme, cathepsin L. Antibodies against purified procathepsin L were raised. These recognized both rat cathepsin L and the purified procathepsin L. To isolate procathepsin L-specific antibodies that did not recognize cathepsin L, sequential affinity chromatography procedures were carried out. Immunoblot analyses showed that the procathepsin L-specific antibodies recognized only procathepsin L, but not cathepsin L.
Ishidoh
K
K
Department of Biochemistry, Juntendo University School of Medicine, Tokyo, Japan.
Takeda-Ezaki
M
M
Kominami
E
E
D016475:3T3 Cells; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D056668:Cathepsin L; D002403:Cathepsins / Q000276:immunology* / Q000302:isolation & purification; D002850:Chromatography, Gel; D003546:Cysteine Endopeptidases; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D004792:Enzyme Precursors / Q000276:immunology* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D051381:Rats
D000906:Antibodies; D004792:Enzyme Precursors; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; C063905:procathepsin L; D056668:Cathepsin L; C534278:Ctsl protein, mouse; C534280:Ctsl protein, rat