A Luminescent Europium Complex for the Sensitive Detection of Proteins and Nucleic Acids Immobilized on Membrane Supports

Lim, Mark J.; Patton, Wayne F.; Lopez, Mary F.; Spofford, K.H.; Shojaee, Negin; Shepro, David

doi:10.1006/abio.1996.9961

Cited by 54 publications

(38 citation statements)

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“…Copper iodide staining is also sensitive but needs complicated steps to prepare reagents [50]. Recently, reversible metal complex stains were introduced, which are compatible with subsequent evaluation [51][52][53].…”

Section: Staining Of Protein On Blotting Membranes With a Visible Dyementioning

confidence: 99%

Usefulness of visible dyes for the staining of protein or DNA in electrophoresis

Choi

2004

Electrophoresis

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Since 1993, we have studied visible organic dye stains for protein or DNA to improve methodologies and developed the counterion dye staining method. The method employs two oppositely charged dyes that form an ion-pair complex in the staining solution. The selective binding of free dye to protein or DNA in the staining solution improves detection sensitivity and speed. It is a rapid and sensitive procedure, involving fixing/staining or staining/quick destaining steps that are completed in 1-1.5 h. The lowest detection limits achieved are 4-8 ng of protein on polyacrylamide gels and approximately 10 ng of DNA on agarose gels. The focus of this review is to chronicle the development and current status of the counterion dye staining method for detection of protein or DNA. As an extended application of visible dyes, we also discuss the visible dye staining method for detecting protein on blotting membranes developed in our laboratory.

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Section: Staining Of Protein On Blotting Membranes With a Visible Dyementioning

confidence: 99%

Usefulness of visible dyes for the staining of protein or DNA in electrophoresis

Choi

2004

Electrophoresis

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“…Densitometry of stained proteins, whether within intact gels or after dissolution or transfer from gels, is a more convenient technology that avoids the radioactive hazards and wastes of the short-lived isotopes. Significant progress in this technique is continually reported with recent emphasis in fluorophore "staining" [9][10][11], but it is still limited in sensitivity, precision, dynamic range, and specificity. Protein or peptide mass spectrometry requires the use of internal standards to achieve broad quantitation, despite its exquisite sensitivity using advanced techniques [12][13][14].…”

Section: Quantitation Of Proteins In Gelsmentioning

confidence: 99%

Attomole quantitation of protein separations with accelerator mass spectrometry

et al. 2001

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Quantification of specific proteins depends on separation by chromatography or electrophoresis followed by chemical detection schemes such as staining and fluorophore adhesion. Chemical exchange of short-lived isotopes, particularly sulfur, is also prevalent despite the inconveniences of counting radioactivity. Physical methods based on isotopic and elemental analyses offer highly sensitive protein quantitation that has linear response over wide dynamic ranges and is independent of protein conformation. Accelerator mass spectrometry quantifies long-lived isotopes such as 14C to subattomole sensitivity. We quantified protein interactions with small molecules such as toxins, vitamins, and natural biochemicals at precisions of 1-5%. Micro-proton-induced X-ray emission quantifies elemental abundances in separated metalloprotein samples to nanogram amounts and is capable of quantifying phopsphorylated loci in gels. Accelerator-based quantitation is a possible tool for quantifying the genome translation into proteome.

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“…A readily reversible, luminescent metal chelate stain consisting of europium complexed to bathophenanthroline disulfonate has been shown to be useful for detection of low-nanogram quantities of proteins immobilized on nitrocellulose or PVDF membranes (16,17). The stain, now commercialized as SYPRO Rose protein blot stain (Molecular Probes), is highly resistant to photobleaching and compatible with popular downstream biochemical characterization procedures including immunoblotting, lectin blotting, and mass spectrometry (17).…”

Section: Discussionmentioning

confidence: 99%

“…The stain, now commercialized as SYPRO Rose protein blot stain (Molecular Probes), is highly resistant to photobleaching and compatible with popular downstream biochemical characterization procedures including immunoblotting, lectin blotting, and mass spectrometry (17). The stain can be excited at 302 nm using standard UV epi-illumination and displays emission maxima at 590 and 615 nm.…”

Section: Discussionmentioning

confidence: 99%

“…detection sensitivity. Ferrozine/ferrous, bathophenanthroline disulfonate/europium, amido black, Fast Green, Coomassie blue, and colloidal silver stain are all medium sensitivity protein stains (1,16,17,20). The colloidal silver stain detects carbonic anhydrase and bovine serum albumin, test proteins described in the original publication, with fairly high sensitivity but stains other proteins present in commercially available molecular weight standards with lower sensitivity (20; Fig.…”

Section: Characteristics Of the Sypro Ruby Staining Proceduresmentioning

confidence: 98%

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A Luminescent Ruthenium Complex for Ultrasensitive Detection of Proteins Immobilized on Membrane Supports

Berggren¹,

Steinberg²,

Lauber

et al. 1999

Analytical Biochemistry

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154

115

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A Luminescent Europium Complex for the Sensitive Detection of Proteins and Nucleic Acids Immobilized on Membrane Supports

Cited by 54 publications

References 30 publications

Usefulness of visible dyes for the staining of protein or DNA in electrophoresis

Usefulness of visible dyes for the staining of protein or DNA in electrophoresis

Attomole quantitation of protein separations with accelerator mass spectrometry

A Luminescent Ruthenium Complex for Ultrasensitive Detection of Proteins Immobilized on Membrane Supports

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