Catalyzed reporter deposition, a novel method of signal amplification application to immunoassays

Bobrow, Mark; Harris, Thomas D.; Shaughnessy, Krista J.; Litt, Gerald J.

doi:10.1016/0022-1759(89)90104-x

Cited by 723 publications

(455 citation statements)

References 10 publications

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“…All antibody incubations were performed for 30 min in a humidified chamber at 37°C and slides were washed with 1 × PBS/0.1% Tween-20/5% normal goat serum. When necessary, signals were amplified using CARD (Catalyzed Reporter Deposition: 26 after the last washing step (that follows the avidin-biotin-peroxidase) biotinylated tyramine was applied on the slide, which was afterwards detected by avidin-peroxidase). In all cases the peroxidase activity was visualised with 3,3Ј-diaminobenzidine tetrahydrochloride (DAB, Sigma) and nuclei were counterstained with haematoxylin.…”

Section: Ish On Isolated Nucleimentioning

confidence: 99%

True

1999

Leukemia

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Gastric MALT lymphoma is a distinct entity related to Helicobacter pylori gastritis. Some studies suggest a role for trisomy 3 in the genesis of these lymphomas, but they mainly focused on low-grade MALT lymphoma. Gastric MALT lymphoma, however, comprises a spectrum from low-to high-grade cases. Furthermore, its exact relation to primary diffuse large B cell lymphoma (DLBCL) of the stomach is not clear. We applied in situ hybridisation (ISH) with centromeric probes on 43 samples of 39 patients with primary gastric lymphoma (13 samples with low-grade MALT lymphoma, 25 with high-grade MALT lymphoma and five with DLBCL) to detect numerical aberrations of 10 chromosomes. ISH was performed immunohistochemically on nuclei isolated from paraffin-embedded resection tissue and on whole paraffin sections using immunofluorescence. In six of 13 low-grade MALT lymphomas trisomy was detected (46%) and mostly involved chromosome 3 (33%). In high-grade MALT lymphomas, trisomies were found in 16 of 25 cases (64%), mainly involving chromosomes 12 and 18. Trisomy 3 was present in only 13% of these cases. Of five DLBCL, only one showed trisomy. Nine of the 16 aberrant high-grade MALT lymphomas (56%) showed trisomy of more than one chromosome per case vs two of six for low-grade cases. In lymphomas with separate low-and high-grade tumour components some trisomies were detected in both components, whereas others occurred only in the high-grade tumour cells. This supports the hypothesis that high-grade MALT lymphomas can develop from a low-grade type and that this progression is accompanied by the acquisition of more genetic aberrations. However, trisomy 3 probably does not play a major role in this progression.

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Section: Ish On Isolated Nucleimentioning

confidence: 99%

True

1999

Leukemia

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“…The field has been plagued by controversy mostly due to differences in techniques used by the different groups to follow cell fate as summarized in [21]. In the last decade a new, very sensitive technique became available utilizing tyramide signal amplification [22,23] and its application to immunohistochemistry was reported [1] describing dilutions of primary antibodies for optimal immunohistochemistry [24] as well as its use in dual immunostaining techniques [25]. Since we also noticed very faint green fluorescent cells in our experimental samples we decided to apply this technique to attempt to visualize most of the GFP expressing cells.…”

Section: Introductionmentioning

confidence: 99%

Sensitive detection of GFP utilizing tyramide signal amplification to overcome gene silencing

Tóth

Shahar

Leker

et al. 2007

Experimental Cell Research

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The green fluorescent protein (GFP) is among the most commonly used expression markers in biology. GFP-tagged cells have played a particularly important role in studies of cell lineage. Sensitive detection of GFP is crucially important for such studies to be successful, and problems with detection may account for discrepancies in the literature regarding the possible fate choices of stem cells. Here we describe a very sensitive technique for visualization of GFP. Using it we can detect about 90% of cells of donor origin while we could only see about 50% of these cells when we employ the methods that are in general use in other laboratories. In addition, we provide evidence that some cells permanently silence GFP expression. In the case of the progeny of bone marrow stem cells, it appears that the more distantly related they are to their precursors, the more likely it is that they will turn off the lineage marker.

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“…In order to get a lower limit of detection, many methods have been developed, including three-dimensional (3D) matrixes modification, such as Noble PA group and Bavykin S group developed ployacrylamide for surface modification [11,12], and Gershwin ME group and Dufva M group used agarose for surface coated and polymer [13,14], which could increase the efficiency of biomolecules immobilization. And some groups chose signal amplification technology [15][16][17][18][19], or used more sensitive signal detection and collection apparatus for signal collection. In our study, we developed a method with Tween coated silica particles that makes the high productivity of the particles.…”

Section: Tweenmentioning

confidence: 99%

Tween-modified suspension array for sensitive biomolecular detection

Zhao

Zong

2012

Chin. Sci. Bull.

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Suspension arrays are attracting increasing interest for detecting and quantifying proteins, nucleic acids and other biomolecules. Among various suspension arrays, sillica-based suspension arrays which benefit from low fluorescence background and material stability have been widely used. Sillica-based suspension arrays are often manufactured with the popular aldehyde-aminosilane chemistry for the attachment of a variety of biomolecules. One drawback of this immobilization strategies is the relatively high array particles lost efficiency when washed by centrifugation. Due to this shortcoming, it is low reproductivity and limited in multiplex assay. Herein we report a novel method to fabricate Tween-coated suspension sillica particles, which could achieve goodreproductivity and the low limit of detection. Tween surfactants, each containing hydrophilic ethylene glycol head groups and a hydrophobic alkyl tail, prevent silica particles from being adsorbed onto the centrifuge tube wall and make beads resuspended well. Also, Tween with low fluorescence background could reduce non-specific protein adsorption. Also Tween surfactants are economic and facile agent. In this study, we demonstrate the preparation of the protein array to substantiate the applicability of our approach. Under the optimized experiment conditions, the limit of detection of our Tween-modified particles is as low as 50 pg/mL, which is sufficiently low for the current methods. We believe that the proposed method could provide a perspective on the improvement of self-encoded silica particle arrays.Tween-20 surfactant, surface modification, suspension array, non-specific adsorption Citation:Zhao Z, Li Z, Li J. Tween-modified suspension array for sensitive biomolecular detection.

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Catalyzed reporter deposition, a novel method of signal amplification application to immunoassays

Cited by 723 publications

References 10 publications

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Sensitive detection of GFP utilizing tyramide signal amplification to overcome gene silencing

Tween-modified suspension array for sensitive biomolecular detection

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