Three-color IHC Staining On Free-floating Sections From Human Brain
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Anne-Sophie Rolland1, Annick Prigent1, Morgan Mathieu2
1Histology platform, Brain and Spine Institute, Hôpital Pitié Salpêtrière, Paris, France
2Enzo Life Sciences, Lausen, Switzerland
Featured Product: MULTIVIEW® PLUS (mouse-HRP/rabbit-AP) IHC Kit (Brown/Green), HIGHDEF® blue IHC chromogen (AP), HIGHDEF® Red IHC Chromogen (HRP), HIGHDEF® yellow IHC chromogen (HRP)
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INTRODUCTION
Immunohistochemistry (IHC) on human brain tissue sections is widely used for diagnosis by anatomopathologists and researchers.
Indeed this technique allows the visualization of specific antigens and thus their distribution into the cells and their compartments.
To handle this type of tissue can be very challenging due to the condition of the brain removal and the quality of the tissue.
There is a growing interest on multiplexing IHC and being able to perform this technique on free-floating human brain sections
as good as formalin-fixed paraffin-embedded sections on slide could be a step forward to analyze larger and thicker specimen
(i.e. 50 μm).
The MULTIVIEW® PLUS (mouse-HRP/rabbit-AP) IHC Kit from Enzo Life Sciences is a non-biotin, one-step detection system suitable
for demonstrating the expression of multiple antigens on tissue sections. It enables faster and more consistent staining procedures
than traditional two-step methods using biotin and avidin/streptavidin conjugates. Indeed, mouse antibodies are directly
tagged with HRP and rabbit antibodies with AP at the same time, all with significantly lower background staining.
The main objective of this study was to validate the use of the MULTIVIEW® PLUS (mouse-HRP/rabbit-AP) IHC Kit and the HIGHDEF
® IHC chromogens to detect multiple staining on human free-floating brain sections. The intensity, sensitivity, and specificity
obtained for all staining demonstrate the capacity of the Enzo Life Sciences kit and their chromogen to achieve multiplexing IHC
on free-floating tissue sections.
Read More about Immunohistochemistry
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