The ultrasensitive silver “protein” stain also detects nanograms of nucleic acids

Somerville, Laura L.; Wang, Kuan

doi:10.1016/0006-291x(81)91487-x

Cited by 86 publications

(23 citation statements)

References 10 publications

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“…In addition, there is a band with the mobility in SDS-PAGE of a 16,000 protein which appears to be RNA as evidenced by its staining with silver, failure to stain with Coomassie Blue, and disappearance with RNAase (27). That this material was nucleic acid was established by phenol extracting the purified Ro/SSA antigen and repeating the SDS-PAGE.…”

Section: Resultsmentioning

confidence: 99%

Molecular properties of the Ro/SSA antigen and enzyme-linked immunosorbent assay for quantitation of antibody.

Yamagata¹,

Harley²,

Reichlin³

1984

J. Clin. Invest.

163

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Section: Resultsmentioning

confidence: 99%

Molecular properties of the Ro/SSA antigen and enzyme-linked immunosorbent assay for quantitation of antibody.

Yamagata¹,

Harley²,

Reichlin³

1984

J. Clin. Invest.

163

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“…Size variants of nebulin in skeletal muscle tissues from human quadriceps (HQ),' snake skeletal muscle (SN), rabbit longissimus dorsi (LD), rabbit psoas (PS), chicken posterior latissimus dorsi (CPLD), and chicken pectoralis (CP) were identified by electrophoresis and Western blots essentially as described (16,23) . For molecular weight calibration, projectin from indirect flight muscle of Lethocerus (6) and human apoliproprotein B were coelectrophoresed as standards.…”

Section: Nebulin Size Variantsmentioning

confidence: 99%

“…Rabbit myofibrillar proteins were first electrophoresed on 2-12 % gradient SDS gels (16) and then transferred to nitrocellulose (NC) paper with a semidry transfer unit (Bio-Rad Laboratories) at 150 mA/gel in KyhseAnderson buffer (8) . The blots were blocked either directly (blocking buffer: 0.1% BSA, 0.2% gelatin, 0.1% Tween-20 in PBS) for 90 min or after boiling for 30 min in H2O to enhance sensitivity of some antibodies (18) .…”

Section: Western Blotsmentioning

confidence: 99%

Nebulin as a length regulator of thin filaments of vertebrate skeletal muscles: correlation of thin filament length, nebulin size, and epitope profile.

Krüger¹,

Wright²,

Wang³

1991

The Journal of Cell Biology

218

175

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Abstract. Nebulin, a family of giant proteins with size-variants from 600 to 900 kD in various skeletal muscles, have been proposed to constitute a set of inextensible filaments anchored at the Z line (Wang, K., and J. Wright . 1988. J. Cell Biol. 107:2199-2212 . This newly discovered filament of the skeletal muscle sarcomere is an attractive candidate for a lengthregulating template of thin filaments . To evaluate this hypothesis, we address the question of coextensiveness of nebulin and the thin filament by searching for a correlation between the size of nebulin variants and the length distribution of the thin filaments in several skeletal muscles . A, positive linear correlation indeed exists for a group of six skeletal muscles that display narrow thin filament length distributions .To examine the molecular and architectural differences of nebulin size-variants, we carried out immunoelectron microscopic studies to map out epitope profiles of nebulin variants in these muscles . For this purpose, a panel of mAbs to distinct nebulin epitopes was produced against rabbit nebulin purified by an imxE lengths ofthick and thin filaments in the sarcomeres ofmost vertebrate skeletal muscles are precisely regulated and are important structural parameters in un derstanding muscle contraction . The mechanisms of such precise length regulation have yet to be elucidated. It is clear from recent studies that although self-assembly of myosin, actin and the contribution of accessory proteins, such as C-protein and tropomyosin, are of major importance in the understanding of the assembly and morphology of myofilaments, these factors by themselves are not sufficient to consistently produce a population of filaments with uniform and defined length (for recent reviews, see 2, 14, 17) . The recent discovery of titin and nebulin, two giant proteins which constitute a filamentous matrix that coexists with thick and thin filaments, has led to the hypothesis that these proteins may A portion of this work has been presented at the 34th Biophysical Society meeting (Kruger, M., J. Wright, and K. Wang . 1990. Biophys. J. 57:554a) .

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“…A modified Gallyas silver stain (SiS) was selected because of its historical perspective in neuroscience and its usefulness in identifying neuronal processes as well as neuropathological hallmarks of neurodegenerative disorders. In addition, SiS has been used to detect minute quantities of RNA 25,26 as well as proteins subjected to gel electrophoresis. A histofluorescent dye, acridine orange (AO), was also employed.…”

mentioning

confidence: 99%

Combined histochemical staining, RNA amplification, regional, and single cell cDNA analysis within the hippocampus

Ginsberg

Che

2004

Laboratory Investigation

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The use of five histochemical stains (cresyl violet, thionin, hematoxylin & eosin, silver stain, and acridine orange) was evaluated in combination with an expression profiling paradigm that included regional and single cell analyses within the hippocampus of post-mortem human brains and adult mice. Adjacent serial sections of human and mouse hippocampus were labeled by histochemistry or neurofilament immunocytochemistry. These tissue sections were used as starting material for regional and single cell microdissection followed by a newly developed RNA amplification procedure (terminal continuation (TC) RNA amplification) and subsequent hybridization to custom-designed cDNA arrays. Results indicated equivalent levels of global hybridization signal intensity and relative expression levels for individual genes for hippocampi stained by cresyl violet, thionin, and hematoxylin & eosin, and neurofilament immunocytochemistry. Moreover, no significant differences existed between the Nissl stains and neurofilament immunocytochemistry for individual CA1 neurons obtained via laser capture microdissection. In contrast, a marked decrement was observed in adjacent hippocampal sections stained for silver stain and acridine orange, both at the level of the regional dissection and at the CA1 neuron population level. Observations made on the cDNA array platform were validated by realtime qPCR using primers directed against b-actin and glyceraldehyde-3 phosphate dehydrogenase. Thus, this report demonstrated the utility of using specific Nissl stains, but not stains that bind RNA species directly, in both human and mouse brain tissues at the regional and cellular level for state-of-the-art molecular fingerprinting studies.

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The ultrasensitive silver “protein” stain also detects nanograms of nucleic acids

Cited by 86 publications

References 10 publications

Molecular properties of the Ro/SSA antigen and enzyme-linked immunosorbent assay for quantitation of antibody.

Molecular properties of the Ro/SSA antigen and enzyme-linked immunosorbent assay for quantitation of antibody.

Nebulin as a length regulator of thin filaments of vertebrate skeletal muscles: correlation of thin filament length, nebulin size, and epitope profile.

Combined histochemical staining, RNA amplification, regional, and single cell cDNA analysis within the hippocampus

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