Colloidal gold immunostaining on deplasticized ultra-thin sections.

Mar, Henderson; Wight, Thomas N.

doi:10.1177/36.11.2844888

Cited by 51 publications

(12 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similarly, Brorson (1997) demonstrated that the immunoreactivity of most antigens is not affected by sodium ethoxide treatment, as ultrathin sections prepared from specimens fixed with formaldehyde and embedded in LR-White, a sodium ethoxide resistant resin, showed no significant difference in immunostaining after sodium ethoxide treatment. Mar and Wight (1988) reported similar results using epon-embedded tissues fixed with a formaldehyde and glutaraldehyde mixture. Whether the great reduction in PCNA-immunostaining by sodium ethoxide treatment is due to the direct modification of the PCNA epitope by ethoxide or the extraction of PCNA because of the destruction of crosslinks between PCNA and neighboring nuclear proteins remains unclear.…”

Section: Discussionsupporting

confidence: 68%

Heat-induced Antigen Retrieval in Conventionally Processed Epon-embedded Specimens

Yamashita

Okada

2014

J Histochem Cytochem.

View full text Add to dashboard Cite

SummaryWe studied the effectiveness of heat-induced antigen retrieval (HIAR) in conventionally processed, epon-embedded specimens and the mechanisms of HIAR in the specimens. Frozen sections were first immunostained to examine the possibility of using HIAR for 18 antigens to avoid the effects of epoxy resin embedment. The antigenicity of 7 out of 18 antigens was retrieved with glutaraldehyde fixation followed by osmium tetroxide treatment whereas none were retrieved with glutaraldehyde fixation without post-osmication. Six antigens also exhibited positive immunostaining in semi-thin epon sections when the sections were deplasticized with sodium ethoxide followed by autoclaving. In the immunoelectron microscopy with the post-embedding method, positive reactions with fine ultrastructures were obtained using HIAR without deplasticization. These results suggested that osmium tetroxide binds to ethylene double bonds (which are introduced into protein crosslinks by glutaraldehyde) and forms an extremely stable resonance interaction with the Schiff bases, thus destabilizing the protein crosslinks. Heating also further degrades these crosslinks. The present study demonstrated that archival epon blocks can be useful resources for immunohistochemical studies for both light and electron microscopy. (J Histochem Cytochem 62:584-597, 2014) Keywords heat-induced antigen retrieval, osmicated and epon-embedded specimen, immunoelectron microscopy, mechanism of antigen retrieval

show abstract

Section: Discussionsupporting

confidence: 68%

Heat-induced Antigen Retrieval in Conventionally Processed Epon-embedded Specimens

Yamashita

Okada

2014

J Histochem Cytochem.

View full text Add to dashboard Cite

show abstract

“…Collagen fibrils present in the adventitia of arterioles in the brain parenchyma ( Figure 2E) also demonstrated positive immunogold labeling of decorin ( Figure 2F). The association of decorin with collagen indicated that the antibody was active and effectively localized to collagen, as others have demonstrated ( Mar and Wight, 1988;Scott and Haigh, 1985;Young, 1985;Scott and Orford, 1981).…”

Section: Nf?s (supporting

confidence: 59%

Peripheral distribution of dermatan sulfate proteoglycans (decorin) in amyloid-containing plaques and their presence in neurofibrillary tangles of Alzheimer's disease.

Snow¹,

Mar²,

Nochlin³

et al. 1992

J Histochem Cytochem.

Self Cite

141

View full text Add to dashboard Cite

We used a polydonal antibody and a mixture of three monoclonal antibodies (MAb), all recognizing the protein core of the s m a l l dermatan sulfate proteoglycan (DSPG) (known as PG-II or decorin) derived from human skin fibroblasts, to immunolocalize this molecule in the characteristic lesions in Alzheimer's brain. All antibodies demonstrated positive decorin immunoStaining in both the amyloid deposits of neuritic plaques (NPs) and the filamentous structures within neurofbrillaty tangles (NFTs). Unlike heparan sulfate proteoglycans (HSPGs), which tend to be evenly distributed throughout NPs containing amyloid fibrils, decorin was pri-

show abstract

“…After fixation, tissue fragments were washed in phosphate buffer, dehydrated through graded alcohols and embedded in Spurr's resin. Semi-thin sections (1.5/~m) were deplasticized using a solution of saturated NaOH in absolute ethanol (sodium ethoxide) for 30 min at room temperature and then processed for immunocytochemical staining [ 16]. Deplasticized sections were immersed in 0.3~o H202 in PBS for 30 min in order to abolish endogenous peroxidase activity.…”

Section: Immunocytolocalization Of Glyoxalase-imentioning

confidence: 99%

Molecular characterization of glyoxalase-I from a higher plant; upregulation by stress

1995

View full text Add to dashboard Cite

A cDNA, GLX1, encoding glyoxalase-I was isolated by differential screening of salt-induced genes in tomato. Glyoxalases-I and -II are ubiquitous enzymes whose functions are not clearly understood. They may serve to detoxify methylglyoxal produced from triosephosphates in all cells. The protein encoded by GLX1 shared 49.4% and 58.5% identity with glyoxalase-I isolated from bacteria and human, respectively. Furthermore, yeast cells expressing GLX1 showed a glyoxalase-I specific activity 20-fold higher than non-transformed cells. Both GLX1 mRNA and glyoxalase-I polypeptide levels increased 2- to 3-fold in roots, stems and leaves of plants treated with either NaCl, mannitol, or abscisic acid. Immunohistochemical localization indicated that glyoxalase-I was expressed in all cell types, with preferential accumulation in phloem sieve elements. This expression pattern was not appreciably altered by salt-stress. We suggest that the increased expression of glyoxalase-I may be linked to a higher demand for ATP generation and to enhanced glycolysis in salt-stressed plants.

show abstract

Colloidal gold immunostaining on deplasticized ultra-thin sections.

Cited by 51 publications

References 0 publications

Heat-induced Antigen Retrieval in Conventionally Processed Epon-embedded Specimens

Heat-induced Antigen Retrieval in Conventionally Processed Epon-embedded Specimens

Peripheral distribution of dermatan sulfate proteoglycans (decorin) in amyloid-containing plaques and their presence in neurofibrillary tangles of Alzheimer's disease.

Molecular characterization of glyoxalase-I from a higher plant; upregulation by stress

Contact Info

Product

Resources

About