PMID: 3772110Oct 23, 1986Paper

S-phase detection with an antibody to bromodeoxyuridine. Role of DNase pretreatment

Journal of Immunological Methods
N J GonchoroffM R Loken

Abstract

We have previously described a monoclonal antibody (BU-1) to 5-bromo-2-deoxyuridine (BrdUrd) that is useful for measurement of cell cycle S-phase. BU-1 hybridoma supernatant reacted with incorporated BrdUrd after the cells had been ethanol fixed; without a requirement for acid or base denaturation. We have found that this reactivity is lost if purified antibody is used, if the culture supernatants are heated, or if a mycoplasma-free hybridoma line is isolated. The supernatant contained endogenous DNase activity that was a result of mycoplasma infection of the cell line. This DNase activity was required for staining the cells with BU-1 in the absence of other denaturation steps. The endogenous DNase could be substituted for by the addition of bovine pancreatic DNase I. The disruption of the double stranded DNA structure with an enzyme rather than with harsh chemical or heat treatments does not affect protein structure or cellular morphology and allows the detection of incorporated BrdUrd of morphologic or antigenic cell subsets. DNase pre-treatment may also be useful for detection of other 'hidden' DNA antigens.

Citations

Nov 1, 1993·Steroids·S SuzukiH Nagasawa
Apr 6, 1988·Journal of Immunological Methods·R Van Furth, T L Van Zwet
Sep 1, 1990·New Zealand Veterinary Journal·S A Holle, M J Birtles
Jan 1, 1993·Cell and Tissue Research·J ArolaA I Kahri
Feb 7, 2015·Journal of Immunotoxicology·Amy PoppleIan Kimber
Jul 29, 2009·Connective Tissue Research·Antonia Icaro CornagliaVittorio Necchi
Jan 22, 2005·Wound Repair and Regeneration : Official Publication of the Wound Healing Society [and] the European Tissue Repair Society·Eric B TredgetAziz Ghahary
Feb 1, 1993·Pathology, Research and Practice·Y HikichiE Sugimoto
Jun 4, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Paolo CappellaJürgen Moll
Jul 19, 2005·Brain Research. Brain Research Protocols·Jorge ValeroJosé R Alonso
May 17, 2002·The Journal of Veterinary Medical Science·Maki MotobuYoshikazu Hirota
Mar 21, 1998·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·J L De GodoyJ E Cardoso
Jun 1, 1996·The British Journal of Surgery·R GillilandP W Hamilton
Nov 1, 1995·The Journal of Pathology·T Matysiak-BudnikM Salaspuro
Jan 1, 1993·European Journal of Immunology·E Paramithiotis, M J Ratcliffe
Jan 1, 1992·Cytometry·P GerlyngP O Seglen
Aug 1, 1997·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·P M Bak, R J Panos
Aug 1, 1990·Environmental Health Perspectives·P O Seglen, P Gerlyng

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