Immuno-localization

Immuno localization using polyclonal antibodies raised against AE showed that AE was widely distributed in orange fruit but with more intensive immunological detection in the outer part of the peels e.g. albedo and flavedo and in the segments (juice vesicles). The results indicate that AE is located at the site where the major fraction of pectin is deposited. 

Immuno localizations of AE in sections of orange fruits are shown in Fig. 3. The most intensive depositions of acetyl esterase were found in the outermost parts of the peel (exocarp or outermost albedo and the flavedo) and in the segments (juice vesicles), although quite high levels of acetyl esterase were found in most other tissues as well. The acetyl esterase depositions were all intracellular. 

Figure 3 Immuno localization of acetyl esterase. Sections were incubated with antibodies raised against the acetyl esterase, followed by visualization with alkaline phosphatase conjugated secondary antibodies and staining with Fast Red.

A Overview of the acetyl esterase immuno localizations in the peel (40x) (Ex exocarp, M mesocarp, OC oil cavity). B Immuno localizations of acetyl esterase in the exocarp (Ex) and oil cavity (OC) (294x). The most intensive acetyl esterase depositions are found in the small sized exocarp cells and in the oil cavity. C Immuno control with preimmune serum on the following section used in B (294x). D Immuno localization of acetyl esterase in endocarp (En) and juice vesicle (JV) (94x). Acetyl esterase depositions in the juice vesicles are more intensive than those observed in the endocarp. No acetyl esterase was detected in the innermost cell layer of the endocarp (see arrows). E Immuno localization of acetyl esterase in lamella (L) and juice vesicle (JV) (294x). Acetyl esterase depositions in the juice vesicles are more intensive than in lamella. Acetyl esterase was absent from the outermost cell layer of lamella (see arrows). F Immuno localization of acetyl esterase in core, where intensive acetyl esterase deposition was found in the xylem (94x). 

Ogihara, H., et al. Immuno-localization of H+/peptide cotransporter in rat digestive tract. Biochem. Biophys. Res. Commun. 1996, 220, 848-852. 

The localization of proteins and carbohydrates within cells and tissues with specific antibodies has long been proven to be a valuable technique. Immuno-localization procedures allow one to detect not only well-characterized cellular structures but also provide information about newly characterized proteins and carbohydrates. This chapter will review some of the advantages and drawbacks of common chemical fixation and permeabilization methods used for immuno-localization at the level of light microscopy. 

Tritscher AM, Goldstein JA, Portier CJ, et al. 1992. Dose-response relationships for chronic exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin in a rat tumor promotion model Quantification and immuno-localization of CYP1A1 and CYP1A2 in the liver. Cancer Res 52 3436-3442. 

Gerard C, Martres MP, Lefevre K, et al. Immuno-localization of serotonin 5-HT6 receptor-like material in the rat central nervous system. Brain Res 1997 746 207-219. 

Foster, R. A., Carpenter, E. J., and Bergman, B. (2006). Unicellular symbionts in open ocean dinoflageUates, radiolarians and tintinnids Ultrastructural characterization and immuno-localization of phycoerythrin and nitrogenase. J. Phycol. 42, 453. 

Important interactions of the nervous and immune systems are known to take place and to involve neuropeptides the largest body of available evidence relates to the participation of substance P in these interactions. Most lymphoid tissues are innervated by substance P positive nerves substance P has also been immuno-localized to immune and inflammatory cells. Cells of the immune system express receptors for substance P, and relatively low concentrations of this peptide alter the functional capabilities of these cells. 

Preparation of T. spiralis Specimens for Immuno-Localization Studies of Premature and Mature Muscle Larvae and Intact Nurse Cells This procedure was adapted from that used for C. elegans (Miller and Shakes 1995). 

Loeffel SG, Gillespie GY, Mirmiran SA, et al. Gellular immuno-localization of SlOO protein within fixed tissue sections by monoclonal antibodies. Arch Pathol Lab Med. 1985 109 117-122. 

Our recent research suggests organ-, tissue-, and cell-specific localization of constitutive and induced terpenoid defense pathways in conifers. For example, linalool synthase (PaTPS-Lin) seems to be preferentially expressed in needles of Norway spruce and Sitka spruce with little or no expression in sterns. ft is also likely that expression of PaTPS-Lin in spruce needles is not associated with resin ducts but could reside in other cells involved with induced terpenoid emission. In contrast, we can speculate that most other mono-TPS and di-TPS are associated with epithelial cells of constitutive and induced resin ducts. The possible localization of conifer sesqui-TPS is difficult to predict. Furthermore, the exact spatial and temporal patterns of terpenoid pathway gene expression associated with traumatic resin duct development in the cambium zone and outer xylem remain to be studied at the tissue and cell level. In situ hybridization and immuno-localization of TPS will address these open questions. These methods have worked well in identifying cell type specific gene and protein expression of alkaloid formation in opium poppy Papaver somniferum) As the biochemistry of induced terpene defenses and the development of traumatic resin ducts have been well described in spruce, this system is ideal for future studies of tissue- and cell-specific localization of transcripts and proteins associated with oleoresin defense and induced volatile emissions in conifers. In addition, the advent of laser dissection microscopy techniques presents a fascinating means by which to further address RNA and protein analysis in a tissue-and cell-specific manner. These techniques, when applied to the cambium zone, xylem mother cells, and the epithelial cells that surround traumatic resin ducts, and will allow a temporal and spatial analysis of cellular functions occurring in the traumatic resin response. 

Identification of specific food components such as proteins and polysaccharides is possible by immuno-localization using gold-labeled antibodies (Figure 3). In conjunction with X-ray microanalysis, SEM is used to analyze salt crystals in cheeses, including... 

Recombinant human core 2 p6-GlcNAc-transferase L has been expressed in CHO cells and was immuno-localized to cis to medial Golgi compartments [53], The enzyme has a similar distribution in the Golgi of normal and cancerous human mammary cells [126],... 

In parallel with efforts to identify the AR-inducer in jelly coat, studies have identified a sperm cell surface molecule that binds jelly coat. In preliminary experiments, it was concluded there must be a receptor-like protein on the surface of the acrosome-intact sea urchin sperm, because trypsin treatment of intact sperm blocked induction of the AR by jelly coat. Subsequently the receptor was isolated and shown to be a 210 kDa glycoprotein with the ability to bind jelly coat [16]. A monoclonal antibody to the 210 kDa glycoprotein was shown to induce the AR. Immuno-localization studies showed the 210 kDa glycoprotein was present on the surface of the sperm flagellum, as well as on a thin belt of the membrane surface... 

Takahashi K, Wesselingh SL, Griffin DE, McArthur JC, Johnson RT, Glass JD (1996) Localization of HlV-1 in human brain using polymerase chain reaction/in situ hybridization and immuno-cytochemistry. Ann Neurol 39(6) 705-711... 

The present work reports the purification and characterization of acetyl esterase from orange fruit as well as the in situ localization of the enzyme by immuno histology. 

Hamada, S., Senzaki, K., Hamaguchi-Hamada, K. et al. (1998). Localization of 5-HT2a receptor in rat cerebral cortex and olfactory system revealed by immuno-histochemistry using two antibodies raised in rabbit and chicken. Molec. Brain Res. 54, 199-211. 

Fig. 17.2. (Opposite) Immuno-gold localization of a T. muris-derived IFN-y homologue to the bacillary band and cuticular pore. (A) Transmission electron micrograph of bacillary band showing the pore chamber (PC), pore aperture (PA) and lamellar apparatus (LA) (x22,000). (B) High-power localization of antibody staining (black dots) to the lamellar apparatus (LA) and pore chamber (PC) (x36,000). (Courtesy ofF. Bughdadi.)...

Brownlee, D.J.A., Fairweather, I., Thomdyke, M.C. and Johnston, C.F. (1996b) Cellular and subcellular localization of SALMFamide (Sl)-like immuno-reactivity within the central nervous system of the nematode Ascaris suum (Nematoda, Ascaroidea). Parasitology Research 82, 149-156. 

De Mey, J., Moeremans, M., Ceuens, G., Nuydens, R., and De Brabander, M. (1981) High resolution light and electron microscopic localization of tubulin with the IgS (immuno gold staining) method. Cell Biol. Int. Rep. 5, 889. 

Tetanus toxin fragment C Tobacco leaves Systemic IgG and local IgA responses. Immuno- -genic in mice when administered nasally. 113... 

Fritschy, J. M., Weinmann, 0., Wenzel, A., and Benke, D. (1998) Synapse-specific localization of NMDA- and GABAA-receptor subunits revealed by antigen-retrieval immuno-histochemistry../. Comp. Neurol. 390,194-210. 

Kim YS, GoodellB, Jellison J. Immuno-electron microscopic localization of extracellular metabolites in spruce wood decayed by brown-rot fungus, Postia placenta. International Research Group on Wood Decay, Stockholm, Sweden, 1990 1441. 

Fig. 11.2 Localization of GFP and calsequestrin (CSQ) in neonatal rat cardiac myocytes. Myo cytes were infected with a recombinant adenovirus containing the cDNAs of GFP and CSQ. Both cDNAs were expressed under control of a separate cytomegalovirus promoter. Expression of CSQ and the coexpressed GFP was detected by fluorescence microscopy. Left-. GFP fluorescence (green), middle immunostaining of CSQ (red), right overlay of GFP fluorescence and immunos taining. Nuclei were counterstained with DAPI (blue). Courtesy of Ulrich Gergs...

Roth, J. (1982) The protein A-gold (pAg) technique—a qualitative and quantitative approach for antigen localization on thin sections, in Techniques in Immuno-cytochemistry, vol. 1 (Bullock, G. R. and Petrusz, P., eds.), Academic, New York, pp. 107-154. 

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