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What are the differences between IHC and ISH and how do they help in the fight against cancer?


Immunohistochemistry (IHC) is an invaluable tool for the detection, localization, and quantification of antigens in preserved tissue for research and diagnostic purposes. This method is widely used in diagnostics and research because of its convenience, reliability, and versatility. In IHC, antigen-antibody complexes can be visualized by fluorescence detection. However, they are generally visualized through light microscopy instead using a color signal, a method referred to as chromogenic detection. The advantage of chromogenic IHC over immunofluorescent techniques is the visible morphology of the tissue around the specific antigen by counterstaining with hematoxylin. Enzo Life Sciences provides IHC Kits which take scientists through the entire workflow, from antigen retrieval to visualization. Results of stained IHC markers are reported semi-quantitatively and have important diagnostic and prognostic implications, particularly for differentiation of benign versus malignant lesions.

Mouse anti-CD44 clone SFF-304 and rabbit anti-Ki-67 clone MIB-1 was used with MULTIVIEW® PLUS (mouse-HRP/rabbit-AP) Kit to perform multiplex immunohistochemistry staining with hematoxylin used as a counterstain.

In situ Hybridization

In situ hybridization (ISH) is a unique molecular analysis method because it provides the precise microscopic localization of nucleic acids such as DNA, mRNA and microRNA in metaphase spreads and cell and tissue preparations. The method was first described in August 1969 by two independent research groups and it was among the first in the collection of methods that have since been classified as molecular biology. In principle, ISH involves (a) the fixation and pre-treatment of the cell or tissue samples to optimally preserve the target nucleic acid, (b) hybridization of a labeled probe by complementary base pairing to the target and lastly (c) detection of the label to allow microscopic visualization of the hybrid. Similar to IHC, detection of the probe can be achieved by chromogenic or fluorescent techniques referred to as chromogenic in situ hybridization (ISH) or fluorescence in situ hybridization (FISH), respectively. We provide efficient labeling methods for FISH that are suitable for a wide range of molecular biology and cytogenetics applications. Today, FISH is an especially powerful tool in the molecular analysis of genetic aberrations that may eventually give rise to the development of cancer.

Nick Translation DNA Labeling System 2.0 was used to label BAC DNA probe for TP53 with Orange 552 dUTP and BAC DNA probe for Centromere 17 with Green 496 dUTP. Labelled probes were hybridized to metaphase spreads. (Institut Universitaire du Cancer Toulouse Oncopole)

Applications in Cancer

IHC in Cancer

IHC, the utilization of monoclonal and polyclonal antibodies for the detection of specific antigens in tissue sections, is an extraordinarily powerful tool in the armamentarium for cancer researchers and diagnostic surgical pathologists. They are able to determine the tissue distribution of an antigen of interest in healthy and diseased states using IHC. It is widely used in cancer research and diagnosis because specific tumor antigens are only expressed de novo or up-regulated in certain cancer types. Therefore, IHC plays an important role in research and pathology, particularly in the subspecialties of oncologic pathology, neuropathology, and hematopathology. While basic histologic examination of tissue is still considered a useful and necessary step, IHC can provide greater and more detailed insights. IHC can be used for various applications in cancer research such as the identification and monitoring of tumor markers, the diagnosis of tumors (particularly for those of uncertain origin), the prognosis of tumors, and to predict therapeutic responses in breast and prostate cancer.

Cervical tissue stained with anti-Ki-67 antibody (mouse), then biotinylated anti-mouse secondary antibody, and followed by SAVIEW® PLUS HRP Reagent

ISH in Cancer

FISH is a powerful technique that is mainly used in the detection of chromosomal abnormalities. The high sensitivity and specificity of FISH and the speed with which the assays can be performed have made FISH a pivotal cytogenetic technique that has provided significant advances in both the research and diagnosis of hematological malignancies and solid tumors. From a medical perspective, FISH can be applied to detect genetic aberrations such as gene fusions, aneuploidy, or loss of a chromosome. It can also be used to monitor the progression of an aberration or for the identification of novel oncogenes. Now that the importance of viral infection in the development of certain cancer types has been widely established, ISH also provides a powerful tool for the diagnosis and research of human papillomavirus (HPV) infection-related cancers in particular. As a long time contributor to the HPV market, Enzo Life Sciences developed the first non-radioactive HPV probes for ISH detection in 1986 and still continues to contribute to the field of HPV Research by providing essential tools for monitoring viral load in cell and tissue samples.

Cervical tissue screened with PATHO-GENE® HPV Type 16/18/31/33/51, followed by SAVIEW® PLUS HRP Reagent

The summary table below highlights the products available for IHC, FISH and ISH. Please click on the product of interest for more information or contact our Technical Support Team for further assistance.
Check out our 10 tips for successful Immunohistochemistry!

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