Gentaur Pub

#29689714   // To Up

Production and characterization of anti-human IgG F(ab')2 antibody fragment.

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Zahra Valedkarimi, Hadi Nasiri, Leili Aghebati-Maleki, Jalal Abdolalizadeh, Mojghan Esparvarinha, Jafar Majidi

2720 related Products with: Production and characterization of anti-human IgG F(ab')2 antibody fragment.

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#21620854   2011/05/18 To Up

Detection of 3-chlorinated tyrosine residues in human cells by flow cytometry.

Hypochlorite is a strong oxidant, generated under pathological conditions, with the potency to introduce chlorine atom into a number of molecules. 3-Chloro- and 3,5-dichlorotyrosine are documented to be generated by this oxidant and their elevated levels were found in many diseases. Thus, we decided to check the possibility of use of FITC-conjugated antibodies for flow cytometric detection of 3-chlorotyrosine residues in human cells (A549, MCF-7, HUVEC-ST) exposed to the action of hypochlorite. Additionally, we compared the effects of chlorohydrins and N-chloroamino acids as chlorine donors. Cell fixation and permeabilization was followed by incubation with rabbit polyclonal anti-3-chlorotyrosine primary antibody and subsequent staining with goat anti-rabbit FITC-labeled secondary antibody. For antibody isotypic control, normal rabbit IgG was employed. Hypochlorite appeared to be the most efficient from the chlorocompounds analyzed in chlorotyrozine generation in all cell lines. Statistically significant increase of fluorescence corresponding to the level of 3-chlorotyrosine residues was found in cells treated with hypochlorite even at non-toxic concentrations (<5μM). This effect was not observed in cells exposed to the action of chlorinated amino acids or chlorohydrins. The use of anti-3-chlorotyrosine antibodies in conjunction with fluorophore-conjugated secondary antibodies analysis allows for detection of 3-chlorotyrosine residues by flow cytometry in cells treated with low doses of hypochlorite.
Agnieszka Robaszkiewicz, Grzegorz Bartosz, Miroslaw Soszynski

2823 related Products with: Detection of 3-chlorinated tyrosine residues in human cells by flow cytometry.

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#20807555   2010/08/31 To Up

Sensitive immunoassay of Listeria monocytogenes with highly fluorescent bioconjugated silica nanoparticles probe.

In this paper, a sensitive immunoassay method was proposed for Listeria monocytogenes detection by using highly fluorescent bioconjugated nanoparticles probe. (FITC-IgG)-doped fluorescent silica nanoparticles (fsNPs) firstly were synthesized by a microemulsion method and characterized by TEM and fluorescent spectra. Then the prepared fsNPs were conjugated with polyclonal rabbit anti-L. monocytogenes antibody (pAb) and used as indicator probe. A sandwich-type immune affinity reaction between polyclonal rabbit anti-L. monocytogenes antibody coated onto microplate wells, target bacteria and the fsNPs-antibody conjugates subsequently was conducted to detect target L. monocytogenes and assemble the indicator probe onto the wells. The target L. monocytogenes was measured by the fluorescent signals of the assembled indicator probes. Under the optimal conditions, the calibration graph of fluorescent intensity is proportional to the amount of target bacteria over the range of 50-10,320 CFU/mL with a detection limit of 50 CFU/mL. The proposed method has been successfully applied to detect L. monocytogenes in food samples offering the advantages of sensitivity, simplicity, and stability.
Zhouping Wang, Tingting Miu, Huan Xu, Nuo Duan, Xiaoying Ding, Shuang Li

2128 related Products with: Sensitive immunoassay of Listeria monocytogenes with highly fluorescent bioconjugated silica nanoparticles probe.

96 wells

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#17723519   2006/03/14 To Up

Combination of immunomagnetic separation with flow cytometry for detection of Listeria monocytogenes.

Listeria monocytogenes can grow at the low temperature commonly used in the storage and transportation of food, and the number of cases of food poisoning caused by L. monocytogenes has increased recently in the US and Europe. Several methods of detecting L. monocytogenes cells have been proposed; however, all existing methods require approximately 48 h incubation. In this study, we attempted rapid detection of L. monocytogenes using flow cytometry (FCM). The method is based on measuring the number of L. monocytogenes cells by using a combination of FCM and immunomagnetic separation (IMS). First, polyclonal antibodies (anti-L. monocytogenes rabbit IgG-FITC) conjugated with fluorescein isothiocyanate (FITC) were reacted with L. monocytogenes cells, and then FCM was applied. The cell numbers were determined by FCM using a traditional colony-counting method in the range of 10(4)-10(8) cells ml(-1). Tetrameric antibody complexes (TAC) were used because they can recognize both magnetic and FITC molecules on the FITC-conjugated antibodies. FITC-labeled L. monocytogenes cells were reacted with a secondary antibody (TAC) bound to magnetic beads. Then, IMS was used. The method is suitable for detection in the range of 10(2)-10(8)cells ml(-1). The FCM assay enumerated the cells within 1 min and the total assay time, including sample preparation, was less than 2 h.
Kyoko Hibi, Akihisa Abe, Eiji Ohashi, Kohji Mitsubayashi, Hideki Ushio, Tetsuhito Hayashi, Huifeng Ren, Hideaki Endo

1967 related Products with: Combination of immunomagnetic separation with flow cytometry for detection of Listeria monocytogenes.

1 kit1 kit200 1 kit1 kit1 kit200 1 kit 5 G1 kit

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#16035233   // To Up

Detection of Xylella fastidiosa from resistant and susceptible grapevine by tissue sectioning and membrane entrapment immunofluorescence.

Immunofluorescence detection was performed by tissue sectioning and membrane entrapment of Xylella fastidiosa from the inoculated hybrid selection F8909-08 (Vitis rupestris A. de Serres x V. arizonica/candicans b43-17; resistant) and Chardonnay (susceptible). In both techniques, tissue sections and bacteria-trapped polycarbonate membranes were incubated with specific polyclonal IgG and stained with fluorescein isothiocyanate (FITC)-conjugated IgG from rabbits to X. fastidiosa cells. The stained preparations were observed by fluorescence microscopy. Rapid identification of the bacteria within 3 weeks post inoculation (wpi) was possible in thin cross sections of the petioles, which allowed penetration of the specific antibody. Examination of the bacteria over time was also possible, and allowed observation of bacterial multiplication and invasion of xylem vessels. The membrane entrapment technique was able to isolate bacteria at low concentrations in infected but asymptomatic plants.
Nihal Buzkan, Lászlo Kocsis, M Andrew Walker

1299 related Products with: Detection of Xylella fastidiosa from resistant and susceptible grapevine by tissue sectioning and membrane entrapment immunofluorescence.

100ug100 mg1000 tests100ug100ul25 mg10 mg500 MG25 mg96T50 ug 1000 TESTS/0.65ml

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#15020090   // To Up

Dual enhancement of triple immunofluorescence using two antibodies from the same species.

Triple immunofluorescence method with two mouse monoclonal antibodies and another rabbit polyclonal antibody was established with catalyzed reporter deposition (CARD) amplification on thick floating sections from the rat cerebellum. One of the monoclonal antibodies (anti-calbindin), diluted maximally, probed with anti-mouse IgG-horseradish peroxidase (HRP) and amplified with Cy5-conjugated tyramide, immunolabeled cerebellar Purkinje cells and their arborization. Subsequently, a rabbit polyclonal IgG (anti-glial fibrillary acidic protein (anti-GFAP)), probed with anti-rabbit IgG-HRP, amplified with biotin-tyramide and visualized with fluorescein-isothiocyanate (FITC)-streptavidin, immunolabeled Bergmann's glia. Another mouse monoclonal IgG (anti-SNAP25), probed with anti-mouse IgG-rhodamine without CARD amplification, selectively visualized synaptic sites, because the maximal dilution of the other monoclonal antibody (anti-calbindin) was below the detection threshold of this anti-mouse IgG-rhodamine. Separation of the two signals (calbindin and SNAP25), each detected through mouse monoclonal antibody, was then based on the difference of sensitivity either with or without CARD amplification. Triple immunofluorescence is possible when just one of the three primary antibodies is from different species. Intensification of two of the three signals provides further advantages to examine immunolocalization of multiple epitopes on histological sections.
Ayako Nakamura, Toshiki Uchihara

2744 related Products with: Dual enhancement of triple immunofluorescence using two antibodies from the same species.

50 ug Product tipe: Antib0.1 mg50 ug Product tipe: Antib100 ug Product tipe: Anti50 ug Product tipe: Antib100 ul Product tipe: Anti100 μg20 ug Product tipe: Antib100 1 ml 100ul

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#12730261   2003/04/28 To Up

In situ localization associates biologically active plant natriuretic peptide immuno-analogues with conductive tissue and stomata.

Plant natriuretic peptide immuno-analogues (irPNP) have previously been shown to affect a number of biological processes including stomatal guard cell movements, ion fluxes and osmoticum-dependent water transport. Tissue printing and immunofluorescent labelling techniques have been used here to study the tissue and cellular localization of irPNP in ivy (Hedera helix L.) and potato (Solanum tuberosum L.). Polyclonal antibodies active against human atrial natriuretic peptide (anti-hANP) and antibodies against irPNP from potato (anti-StPNP) were used for immunolabelling. Tissue prints revealed that immunoreactants are concentrated in vascular tissues of leaves, petioles and stems. Phloem-associated cells, xylem cells and parenchymatic xylem cells showed the strongest immunoreaction. Immunofluorescent microscopy with fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG supported this finding and, furthermore, revealed strong labelling to stomatal guard cells and the adjacent apoplastic space as well. Biologically active immunoreactants were also detected in xylem exudates of a soft South African perennial forest sage (Plectranthus ciliatus E. Mey ex Benth.) thus strengthening the evidence for a systemic role of the protein. In summary, in situ cellular localization is consistent with physiological responses elicited by irPNPs reported previously and is indicative of a systemic role in plant homeostasis.
M M Maryani, M V Morse, G Bradley, H R Irving, D M Cahill, C A Gehring

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