PMID: 7015422Mar 1, 1981Paper

Sequential use of the PAP and immunogold staining method for the light microscopical double staining of tissue antigens

Regulatory Peptides
J GuJ M Polak

Abstract

Double immunoperoxidase staining using different couplers can give various combinations of colours on a single tissue section to achieve a comparable picture of different antigens. However, the colour combinations achieved to date are not entirely satisfactory. A double immunostaining procedure is introduced here, combining the peroxidase anti-peroxidase (PAP) and immunogold staining (IGS) methods. The IGS method is a new, simple, sensitive and reliable approach to immunostaining at the light microscopic level. It was carried out in three ways. Firstly, a two-step method was used in which the second layer was goat anti-rabbit IgG absorbed onto gold particles (GAR/Au20). Secondly, a three-step method was employed where the second layer was unlabelled goat anti-rabbit IgG and the third layer was a rabbit antibody to peroxidase absorbed onto the gold particles (RAP/Au20) and acting as a gold-labelled IgG antigen. The third method combined the first two methods using GAR/Au20 as th second layer and RAP/Au20 as the third layer which increased the amount of bound gold and enhanced the red colour, providing a better picture. The use of gold-labelled antibodies in double immunostaining has great potential value for many studies includi...Continue Reading

References

Mar 1, 1975·The Histochemical Journal·A G Pearse, J M Polak
Aug 1, 1974·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·J P PetraliL A Sternberger
Sep 1, 1968·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·P K Nakane
Apr 1, 1966·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·R C Graham, M J Karnovsky
Jul 1, 1980·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·N Van Rooijen

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Citations

Jan 1, 1984·Histochemistry·D R SpringallJ M Polak
Jan 1, 1985·Histochemistry·P M LackieJ M Polak
Jul 1, 1996·Histochemistry and Cell Biology·P M Lackie
Dec 1, 1985·General and Comparative Endocrinology·A P LamersA M Michelakis
Jul 1, 1994·Journal of Neuroscience Methods·R Wegerhoff, O Breidbach
Jan 1, 1981·Peptides·J M Polak, S R Bloom
Jan 7, 2014·Histochemistry and Cell Biology·Jing LiJiang Gu
Jan 1, 1992·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·R Magar, R Larochelle
Apr 1, 2009·Laboratory Investigation; a Journal of Technical Methods and Pathology·Na NiuJiang Gu
Jun 1, 1989·The American Journal of Anatomy·P A In't Veld
Jan 1, 1991·Journal of Tongji Medical University = Tong Ji Yi Ke Da Xue Xue Bao·L Q Xia, Q T Wang
Jan 1, 1985·Virchows Archiv. A, Pathological Anatomy and Histopathology·G W HackerJ M Polak
Jul 21, 2006·The Journal of Physical Chemistry. B·Matthias Koebel, Matthew B Zimmt

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