Citraconic anhydride: a new antigen retrieval solution

Haffajee, Zenobia

doi:10.3109/00313020903434439

Cited by 16 publications

(8 citation statements)

References 13 publications

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“…This was exactly the same case of the French authors when they proved that the fine blue granules in the amoeba were actually Mimiviruses 6 . To enhance the resolution detection of the EM, we used a particular antigen retrieval solution with citraconic anhydride and heat 7, 8 . EM analyses were conducted in two different international centres and 300 micrographs were scrutinized by operators who performed a blinded reading and were also blind to each other.…”

Section: Resultsmentioning

confidence: 99%

Questions on unusual Mimivirus-like structures observed in human cells

et al. 2017

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Background: Mimiviruses or giant viruses that infect amoebas have the ability to retain the Gram stain, which is usually used to colour bacteria. There is some evidence suggesting that Mimiviruses can also infect human cells. Guided by these premises, we performed a routine Gram stain on a variety of human specimens to see if we could detect the same Gram positive blue granules that identify Mimiviruses in the amoebas. Methods: We analysed 24 different human specimens (liver, brain, kidney, lymph node and ovary) using Gram stain histochemistry, electron microscopy immunogold, high resolution mass spectrometry and protein identification. Results: We detected in the human cells Gram positive granules that were distinct from bacteria. The fine blue granules displayed the same pattern of the Gram positive granules that diagnose Mimiviruses in the cytoplasm of the amoebas. Electron microscopy confirmed the presence of human Mimiviruses-like structures and mass spectrometry identified histone H4 peptides, which had the same footprints as giant viruses. However, some differences were noted: the Mimivirus-like structures identified in the human cells were ubiquitous and manifested a distinct mammalian retroviral antigenicity. Conclusions: Our main hypotheses are that the structures could be either giant viruses having a retroviral antigenicity or ancestral cellular components having a viral origin. However, other possible alternatives have been proposed to explain the nature and function of the newly identified structures.

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Section: Resultsmentioning

confidence: 99%

Questions on unusual Mimivirus-like structures observed in human cells

et al. 2017

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“…Other immunohistochemistry studies have reported that paraffin-embedded sections following microwave retrieval can preserve tissue structure and protect the antigens. [10][11][12][13][14][15][16] Use of conventional antigen retrieval reagents allows visualization of an antibody with a weak signal, thereby increasing the sensitivity of staining. [17][18][19] Immunohistochemistry of paraffin-embedded sections following microwave retrieval was reported with satisfactory findings, improving the specific localization of coexpressed antigens, which is very useful for observing correlations between tissue structure and deposition of immune complexes.…”

Section: Discussionmentioning

confidence: 99%

Immunofluorescence With Dual Microwave Retrieval of Paraffin-Embedded Sections in the Assessment of Human Renal Biopsy Specimens

Shu

Cheng

Zhang

et al. 2013

American Journal of Clinical Pathology

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Immunofluorescence of frozen tissue sections (IF-F) is a classic technique for renal immunopathologic examination. However, it has certain disadvantages, such as diffuse antigen distribution and few or even no glomeruli in the section. We developed a new technique of immunofluorescence staining using dual microwave retrieval in paraffin-embedded renal tissue sections (IF-DMP) and compared IF-DMP with IF-F in 406 renal biopsy samples. IF-DMP detected significantly more glomeruli than did IF-F (P< .001). There was no significant difference for the specificity and sensitivity in the detection of immunoglobulins, complements, κ, and λ between IF-F and IF-DMP. Concordant observations were 98% for all immunofluorescence, complements, κ, and λ staining and 100% for immunoglobulin staining. Both techniques were completely accurate in confirming diagnoses of various glomerular diseases. IF-DMP provided clearer images of tissue structure and more precise localization of antigens, and it is a suitable alternative for traditional IF-F in clinical renal immunopathologic diagnosis.

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“…Tissue sections (4 µm) were de-waxed and rehydrated (Blaschitz et al 2000 ). Antigen retrieval was done with 0.05% (v/v) citraconic anhydride solution, pH 7.4, for 20 min (Leong and Haffajee 2010 ). Sections were incubated with blocking buffer (5% (v/v) goat serum (Jackson ImmunoResearch Laboratories; 005-000-121) in PBS containing 0.05% (w/v) saponin (Sigma; SAE0073) for 1 h at room temperature.…”

Section: Methodsmentioning

confidence: 99%

In vitro function and in situ localization of Multidrug Resistance-associated Protein (MRP)1 (ABCC1) suggest a protective role against methyl mercury-induced oxidative stress in the human placenta

et al. 2020

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Methyl mercury (MeHg) is an organic highly toxic compound that is transported efficiently via the human placenta. Our previous data suggest that MeHg is taken up into placental cells by amino acid transporters while mercury export from placental cells mainly involves ATP binding cassette (ABC) transporters. We hypothesized that the ABC transporter multidrug resistance-associated protein (MRP)1 (ABCC1) plays an essential role in mercury export from the human placenta. Transwell transport studies with MRP1-overexpressing Madin-Darby Canine Kidney (MDCK)II cells confirmed the function of MRP1 in polarized mercury efflux. Consistent with this, siRNA-mediated MRP1 gene knockdown in the human placental cell line HTR-8/SVneo resulted in intracellular mercury accumulation, which was associated with reduced cell viability, accompanied by increased cytotoxicity, apoptosis, and oxidative stress as determined via the glutathione (GSH) status. In addition, the many sources claiming different localization of MRP1 in the placenta required a re-evaluation of its localization in placental tissue sections by immunofluorescence microscopy using an MRP1-specific antibody that was validated in-house. Taken together, our results show that (1) MRP1 preferentially mediates apical-to-basolateral mercury transport in epithelial cells, (2) MRP1 regulates the GSH status of placental cells, (3) MRP1 function has a decisive influence on the viability of placental cells exposed to low MeHg concentrations, and (4) the in situ localization of MRP1 corresponds to mercury transport from maternal circulation to the placenta and fetus. We conclude that MRP1 protects placental cells from MeHg-induced oxidative stress by exporting the toxic metal and by maintaining the placental cells' GSH status in equilibrium.

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Citraconic anhydride: a new antigen retrieval solution

Cited by 16 publications

References 13 publications

Questions on unusual Mimivirus-like structures observed in human cells

Questions on unusual Mimivirus-like structures observed in human cells

Immunofluorescence With Dual Microwave Retrieval of Paraffin-Embedded Sections in the Assessment of Human Renal Biopsy Specimens

In vitro function and in situ localization of Multidrug Resistance-associated Protein (MRP)1 (ABCC1) suggest a protective role against methyl mercury-induced oxidative stress in the human placenta

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