Prevention of false positive binding during immunofluorescence of Staphylococcus aureus infected tissue biopsies
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- 作者：Neil C.-W. Tan ; Hai Tran ; Eugene Roscioli ; Peter John Wormald ; Sarah Vreugde
- 关键词：F(ab&prime ; ) ; fragment antigen-binding region of antibody ; Fc ; fragment crystallizable region of antibody ; MSCRAM ; microbial surface components recognising adhesive matrix molecules ; TBS-T ; Tris-buffered saline and 0.05 % tween-20 buffer ; SFB ; serum free p
- 刊名：Journal of Immunological Methods
- 出版年：2012
- 出版时间：31 October, 2012
- 年：2012
- 卷：384
- 期：1-2
- 页码：111-117
- 全文大小：847 K
文摘
Immunofluorescence is a fundamental tool used to analyse tissue and cell samples with a wide variety of available antibodies targeting specific proteins or molecules. Staphylococcal surface protein A is used both in clinical, research and industrial settings for its ability to bind mammalian immunoglobulin G. Spurious binding between protein A and IgG antibodies can lead to false-positive fluorescence and misleading results. Here we demonstrate this occurring in formalin-fixed patient samples that harbour Staphylococcus aureus infection, and characterise methods to overcome this issue. Specifically the use of F(ab¡ä) fragment antibodies or blocking with human IgG is shown to prevent antibody-protein A interaction in formalin-fixed S. aureus smears, biopsies obtained from infected patients, and experimentally infected tissue samples.