Staining of proteins on SDS polyacrylamide gels and on nitrocellulose membranes by Alta, a colour used as a cosmetic

Pal, Jayanta K.; Godbole, Dhanashri; Sharma, Kiran K.

doi:10.1016/j.jbbm.2004.06.008

Cited by 21 publications

(6 citation statements)

References 6 publications

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“…Pal et al (23) describe the use of Alta, a pre-existing scarlet-red stain used for cosmetic purposes, for staining gels and nitrocellulose membranes during western blot analysis. Alta is made up of 0.8% Crocein scarlet (brilliant Crocein) and 0.2 % Rhodamine B.…”

Section: Membranes By Alta a Color Used As A Cosmeticmentioning

confidence: 99%

Other Notable Methods of Membrane Protein Detection: A Brief Review

Kurien

Scofield

2015

Methods in Molecular Biology

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Several techniques have been employed to detect proteins on membranes. These include the use of quantum dot luminescent labels, oxyblot immunochemical detection, polymer immunocomplexes, "coupled" probing approach, in situ renaturation of proteins for detecting enzyme activities in crude or purified preparations, immunochromatographic assay, western-phosphatase assay, and use of Congo red dye (a cosmetic color named Alta), Pro-Q Emerald 488 dye, or amine-reactive dye in combination with alkaline phosphatase- and horseradish peroxidase-antibody conjugates for the simultaneous trichromatic fluorescence detection of proteins. Several methods have been used to improve the detection of proteins on membranes, including glutaraldehyde treatment of nitrocellulose blots, elimination of keratin artifacts in immunoblots probed with polyclonal antibodies, and washing of immunoblots with excessive water and manipulation of Tween-20 in wash buffer. These methods are briefly reviewed in this chapter.

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Section: Membranes By Alta a Color Used As A Cosmeticmentioning

confidence: 99%

Other Notable Methods of Membrane Protein Detection: A Brief Review

Kurien

Scofield

2015

Methods in Molecular Biology

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“…Alternate densitometric stains have not gained popularity within the proteomic research community because these stains have been unable to compete with the performance of fluorescent staining and imaging. Also, most of these alternate densitometric stains have, at best, had limited characterization or have similar detection sensitivity to cCBB and hence have not been pursued for further application in proteomics [54,[117][118][119][120][121][122][123][124][125][126].…”

Section: Additional Densitometric Stainsmentioning

confidence: 99%

Quantitative proteomics: assessing the spectrum of in-gel protein detection methods

2010

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Proteomics research relies heavily on visualization methods for detection of proteins separated by polyacrylamide gel electrophoresis. Commonly used staining approaches involve colorimetric dyes such as Coomassie Brilliant Blue, fluorescent dyes including Sypro Ruby, newly developed reactive fluorophores, as well as a plethora of others. The most desired characteristic in selecting one stain over another is sensitivity, but this is far from the only important parameter. This review evaluates protein detection methods in terms of their quantitative attributes, including limit of detection (i.e., sensitivity), linear dynamic range, inter-protein variability, capacity for spot detection after 2D gel electrophoresis, and compatibility with subsequent mass spectrometric analyses. Unfortunately, many of these quantitative criteria are not routinely or consistently addressed by most of the studies published to date. We would urge more rigorous routine characterization of stains and detection methodologies as a critical approach to systematically improving these critically important tools for quantitative proteomics. In addition, substantial improvements in detection technology, particularly over the last decade or so, emphasize the need to consider renewed characterization of existing stains; the quantitative stains we need, or at least the chemistries required for their future development, may well already exist.

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“…A single armed band was detected by SDS-PAGE (Fig. 7), Staining and destaining were done as described [20].…”

Section: Resultsmentioning

confidence: 99%

Expression of porcine parvovirus VP2 gene requires codon optimized E. coli cells

Cui

2009

Virus Genes

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Porcine parvovirus (PPV) is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. The gene coding for the capsid protein VP2 of PPV was amplified and inserted into the plasmid pET-32a (+), which was then used to transform Escherichia coli Rosetta, the capsid protein of PPV was fused to a polyhistidine tag, and the position of the affinity tag is in N-terminus. VP2 was expressed using different expression host bacteria, including E. coli BL21, and Rosetta, and different plasmid vectors, including pET-30a (+), pET-32a (+), and pGEX-6p-1. After selection, only the fusion protein inserted into pET-32a (+) was expressed well in E. coli Rosetta. The recombinant bacterium produced high quantities of the fusion protein VP2, about 8% in total. The expressed VP2 was antigenically similar to the native capsid protein according to a Western blot assay performed with polyclonal antibodies obtained from pigs vaccinated with PPV. A simple, easily commercialized procedure was used to purify this protein. This study provides a foundation for the application of VP2 protein in the clinical diagnosis of PPV and in the vaccination against PPV.

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Staining of proteins on SDS polyacrylamide gels and on nitrocellulose membranes by Alta, a colour used as a cosmetic

Cited by 21 publications

References 6 publications

Other Notable Methods of Membrane Protein Detection: A Brief Review

Other Notable Methods of Membrane Protein Detection: A Brief Review

Quantitative proteomics: assessing the spectrum of in-gel protein detection methods

Expression of porcine parvovirus VP2 gene requires codon optimized E. coli cells

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