Polyclonal antibodies against RNase L. Subcellular localization of this enzyme in mouse cells.
RNase L activated by 2-5A (a series of 2'-5'-linked adenylic oligoribonucleotides) is a key enzyme of the interferon system. To study RNase L (endonuclease L) in intact cells independently of intracellular 2-5A and of its activity, we have developed polyclonal antibodies against RNase L. RNase L from mouse spleen was purified on a column of 2-5A-Sepharose and used to immunize rabbits in co-injection with polyadenylic-polyuridylic acid as adjuvant. Antibodies were purified by chromatography on Affi-Gel blue and 2-5A-Sepharose-immobilized RNase L. These polyclonal antibodies immunoprecipitate the 80- and 40-kDa forms of RNase L in mouse spleen. In Western blot, only the 80-kDa form of RNase L is recognized by these antibodies. These purified antibodies were used to localize RNase L in the cytoplasm of intact mouse NIH 3T3 cells by immunofluorescence. The cytoplasmic localization of RNase L was confirmed by its 2-5A binding activity after cellular fractionation.
Salehzada
T
T
Laboratoire de Biochimie des Protéines, Unité de Recherche Associée Centre National de la Recherche Scientifique 1191, Université de Montpellier II Sciences et Techniques du Languedoc, France.
Silhol
M
M
Lebleu
B
B
Bisbal
C
C
D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D002458:Cell Fractionation; D002460:Cell Line; D004722:Endoribonucleases / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D007372:Interferons / Q000494:pharmacology; D051379:Mice; D011233:Precipitin Tests; D011817:Rabbits; D012270:Ribosomes / Q000378:metabolism; D013154:Spleen / Q000201:enzymology
D000906:Antibodies; D007372:Interferons; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease