Immunohistochemistry

This technique is a semiquantitative means of determining the presence of a protein within a cell or tissue section, its abundance, and its subcellular localization. This approach could be used in toxicology, for example, to determine if, upon treating cells with a specific toxicant, a receptor to which the toxicant binds localizes from the cell cytoplasm to the nucleus. 

The general approach used to assess the intracellular localization of a protein within a cell is as follows. 

Note Avoid exposing the cel Is to I ight as much as possible from step 15 onwards. For suspension cells carry out the staining protocol in an eppendorf tube till step 10. To adhere the cell 

Immunohistochemistry is used for detecting the constituents of tissue by using specific antibody tagged to a visibie iabei which will bind to the antigen inside the tissue. The tissues are paraffinised, frozen or embedded in resin. 

Dissect the tissue out and transfer into 2% paraformaldehyde solution. 

Incubate in fixative for 30 min or overnight depending on the size of the tissue. 

Section the tissue with the thickness of less than 10 ixmeters and fix on slide. 

Phenotypic and Genotypic Diagnosis of Malignancies. Muin S.A. TufFaha 

Copyright 2008 WILEY-VCH Verlag GmbH Co. KGaA, Weinheim 

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ADCC. Cetuximab is approved for treatment of metastatic colorectal cancer (CRC) and squamous cell carcinoma of the head and neck (SCCHN). Interestingly, an adverse event, acneiform rash seems to correlate with a better response to cetuximab, while there is no such correlation with expression levels of EGFR assessed by immunohistochemistry. Further side effects are rare infusion reactions and hypomagnesia. Two other anti-EGFR antibodies approved for clinical use are the fully human antibody panitumumab (Vectibix)... 

Mastocytosis is recognized in most patients because of the presence of characteristic cutaneous lesions [10]. A positive Darier s sign and/or histological examination of the skin using metachromatic stains, or by immunohistochemistry using antibodies to mast cell tryptase, helps confirm the diagnosis of cutaneous disease. 

Osawa M, Satoh F, Horiuchi H, Tian W, Kugota N, Hasegawa I Postmortem diagnosis of fatal anaphylaxis during intravenous administration of therapeutic and diagnostic agents evaluation of clinical laboratory parameters and immunohistochemistry 25 in three cases. Leg Med (Tokyo) 2008 10 143-147. 

Early neuronal damage detected by immunohistochemistry has been confirmed by brain magnetic resonance spectroscopy (MRS) analysis of the neuronal marker N-acetylaspartate (NAA) (commonly expressed as an NAA/creatine ratio NAA/ Cr) in the acute and chronic phases of infection in SIV-infected macaques (Fuller et al. 2004 Greco et al. 2004 Lentz et al. 2005 Lentz et al. 2008 Williams et al. 2005). In a macaque model involving CD8h- T lymphocyte depletion along with SIV inoculation, Williams et al. (2005) demonstrated a reduction in NAA/Cr in the... 

Bagasra O, Lavi E, Bobroski L, Khafili K, Pestaner JP, Tawadros R, Pomerantz RJ (1996) CeUular reservoirs of HIV-1 in the central nervous system of infected individuals identification by the combination of in situ polymerase chain reaction and immunohistochemistry. Aids 10(6) 573-585... 

It was shown by immunohistochemistry on post-mortem sections, that CXCR2 was expressed in neurons of various regions of the human adult nervous system including the hippocampus, cerebellum, pontine nuclei and spinal cord. The labeling... 

IGSS Immuno-gold silver stain IHC Immunohistochemistry IHES Idiopathic hypereosinophilic syndrome... 

Steinbusch, H.W.M. Verhofstad, A.A.J. and Joosten, H.W.J. Localization of serotonin in the central nervous system by immunohistochemistry Description of a specific and sensitive technique and some applications. Neuroscience 3 811-819, 1978. 

Significant additional interest in the potential role of oxytocin in labour developed from 1993, when it was reported for the first time that oxytocin, which had previously been thought to only be produced in the hypothalamus, may also be synthesised locally in the uterus as indicated by the presence of mRNA encoding for oxytocin [11]. This observation has recently been confirmed by using immunohistochemistry and mass spectrometry to confirm the presence of local oxytocin peptide in the uterus. These studies localised oxytocin to the chorionic trophoblast, decidual stroma and glandular epithelium, and demonstrated that local uterine concentrations of oxytocin increased with the onset of labour [12]. 

Hematopathology evaluation of biopsy specimen-morphologic inspection, immunohistochemistry for cell-surface antigens to characterize lymphoma cells, cytogenetic analysis... 

FIGURE 24.3 Distribution of CBP. (a) Analysis of tissue distribution by Western blotting, (b) Immunohistochemistry of the midgut epithelium. CBP was not detected in the goblet cells (G) of the midgut epithelium, which are thought to be involved in ion transport. 

Takeda, N., Inagaki, S., Taguchi, Y. et al. (1984). Origins of histamine-containing fibers in the cerebral cortex of rats studied by immunohistochemistry with histidine decarboxylase as a marker and transection. Brain Res. 323, 55-63. 

Kalia, M., O Callaghan, J.R, Miller, D.B., and Kramer, M., Comparative study of fluoxetine, sibutra-mine, sertraline and dexfenfluramine on the morphology of serotonergic nerve terminals using serotonin immunohistochemistry, Brain Res. 858(1), 92-105, 2000. 

Sanmartin, M.L., Iglesias, R., Santamarina, M.T., Leiro,J. and Ubeira, F.M. (1991) Anatomical location of phosphorylcholine and other antigens on encysted TrichineUa using immunohistochemistry followed by Wheadey s trichrome stain. Parasitology Research 77, 301-306. 

MacDonald, T.T., Choy, M.Y., Spencer, J., Richman, P.I., Diss, T., Hanchard, B., Venugopal, S., Bundy, D.A. and Cooper, E.S. (1991) Histopathology and immunohistochemistry of the caecum in children with the Trichuris dysentery syndrome. Journal of Clinical Pathology 44, 194-199. 

Fig. 20.3 Chronic lithium increases hippocampal neurogenesis. C57BL/6 mice were treated with lithium for 14 days, and then received once daily BrdU injections for 12 consecutive days while lithium treatment continued. 24 hours after the last injection, the brains were processed for BrdU immunohistochemistry.

Antibodies raised against the Ap peptide were subsequently used to identify more diffuse types of Ap-containing deposits in AD brain by immunohistochemistry. Diffuse Ap peptide deposits lack the... 

Antigen Retrieval Immunohistochemistry Based Research and Diagnostics, Edited by Shan-Rong Shi and Clive R. Taylor Copyright 2010 John Wiley Sons, Inc. 

Taylor CR, Levenson RM. Quantification of immunohistochemistry—issues concerning methods, utility and semiquantitative assessment II. Histopathology 2006 49 411 124. 

Taylor CR. Standardization in immunohistochemistry the role of antigen retrieval in molecular morphology. Biotech. Histochem. 2006 81 3-12. 

Shi SR, Cote RJ, Taylor CR. Antigen retrieval immunohistochemistry past, present, and future./. Histochem. Cytochem. 1997 45 327-343. 

Shi S-R, Cote RJ, Chaiwun B, et al. Standardization of immunohistochemistry based on antigen retrieval technique for routine formalin-fixed tissue sections. Appl. Immunohistochem. 1998 6 89-96. 

Shi S-R, Cote RJ, Taylor CR. Standardization and further development of antigen retrieval immunohistochemistry strategies and future goals. I. Histotechnol. 1999 22 177-192. 

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