The effect of various stains on quantitative agarose-gel electrophoresis: Evaluation of the results with the aid of an analog computer

Siegert, Martín J.; Schützler, G.; Jarofke, R.

doi:10.1016/0009-8981(76)90143-1

Cited by 5 publications

(2 citation statements)

References 8 publications

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“…Cold Springs Harbor recommends 0.5% (w/v) Ponceau S in 1% (v/v) acetic acid [16], EncorBio advises 0.25% (w/v) Ponceau S in 40% methanol and 15% acetic acid [17], while Abcam suggests 0.02% (w/v) Ponceau S in 0.3% (w/v) TCA and 0.3% (w/v) sulfosalicyclic acid [18]. To further complicate the use of Ponceau S, scientists have also used other compositions of Ponceau S stain including 0.6% (w/v) Ponceau S in 3% (w/v) TCA [19]. The methods recommended to stain and destain membranes with Ponceau S stain is also variable ( Table 1).…”

Section: Resultsmentioning

confidence: 99%

Ponceau S waste: Ponceau S staining for total protein normalization

Sander

Wallace

Plouse

et al. 2019

Analytical Biochemistry

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Normalization of Western blotting data is a critical step that is needed to reduce errors caused by unequal sample loading across lanes in a gel, inconsistent sample preparation, and variations due to experimental errors. Several papers have suggested that total protein normalization may be better than housekeeping protein normalization for Western blotting normalization. Ponceau S is the most commonly used stain for total protein normalization. A review of the literature and commercial websites suggest a multitude of Ponceau S staining protocols for total protein staining of blots. In this study, we explored which Ponceau S staining protocol would result in the highest sensitivity of protein band detection. Unexpectedly, we found that irrespective of the Ponceau S concentration (between 0.001-2% (w/v)), acid concentration, and acid type (acetic acid, trichloroacetic acid and/or sulfosalicylic acid), the sensitivity of protein detection remained constant. The most commonly used concentration of Ponceau S is 0.1%, while 0.001% (100-fold less) Ponceau S resulted in the same sensitivity of protein band detection. We suggest the use of the relatively inexpensive 0.01% Ponceau S in 1% acetic acid stain for total protein normalization as it is as effective as all the expensive formulations that are currently used.

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Section: Resultsmentioning

confidence: 99%

Ponceau S waste: Ponceau S staining for total protein normalization

Sander

Wallace

Plouse

et al. 2019

Analytical Biochemistry

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“…For our tests, we made use of Amido Black 10 B, after it had proved that, contrary to the results published by other authors (28,29), this easily handled stain, although chromatogjaphically unhomogenous, is quite suitable for quantitative determination (30).…”

Section: Discussionmentioning

confidence: 95%

Agarose Gel Electrophoresis of Cerebrospinal Fluid Proteins and Analysis of the Pherogram Profiles by Analog Computer

Siegert¹,

Siemes²

1977

Clinical Chemistry and Laboratory Medicine

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A micro-method of agarose gel electrophoresis requiring I-3 ml of cerebrospinal fluid for quantitative analysis of cerebrospinal fluid proteins is described. After concentration of CSF to about 50 μΐ by ultrafiltration and refractometric determination of protein, approximately 20-40 ^g of total protein are used for electrophoresis. Photometric scanning of the electrophoretic pattern at two different wavelengths permits quantitative evaluation. The pherograms are analysed by means of a modified DU-PONT analog computer. Factors which influence quantitative electrophoresis are examined. In cerebrospinal fluid of normal children 15 protein fractions are demonstrated: 2 prealbumins, albumin, 5α-, 3/3-and 47-globulins. Agarosegelelektrophorese von Liquorproteinen und Analyse der Pherogrammprofile mit dem AnalogrechnerZusammenfassung: Es wird ein Mikroverfahren zur quantitativen Elektrophorese der Liquorproteine in Agarosegel beschrieben. Zur Analyse sind 1-3 ml Liquor cerebrospinalis erforderlich. Nach der Einengung mittels Ultrafiltration auf 50 μΐ und der refraktometrischen Eiwei bestimmung im Konzentrat werden ca. 20-40 ^g Gesamtprotein zur Elektrophorese eingesetzt. Die Photometric des Elektrophoresemusters bei 2 verschiedenen Wellenl ngen erlaubt eine quantitative Auswertung. Die Analyse der Pherogramme erfolgt mit einem modifizierten DU-PONT Analogrechner. Es werden Faktoren untersucht, welche die quantitative Elektrophorese beeinflussen. Im Liquor cerebrpspinalis normaler Kinder k nnen 15 Proteinfr kti nen nachgewiesen werden: 2 Pr albumine, Albumin, 5α-, 3 β-und 4yrGlobuline.

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Derivative electrophoretograms and their applications

Okamura

1979

Clinica Chimica Acta

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The effect of various stains on quantitative agarose-gel electrophoresis: Evaluation of the results with the aid of an analog computer

Cited by 5 publications

References 8 publications

Ponceau S waste: Ponceau S staining for total protein normalization

Ponceau S waste: Ponceau S staining for total protein normalization

Agarose Gel Electrophoresis of Cerebrospinal Fluid Proteins and Analysis of the Pherogram Profiles by Analog Computer

Derivative electrophoretograms and their applications

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