Use of 5-(4-dimethylaminobenzylidene)rhodanine in quantitating silver grains eluted from autoradiograms of biological material

Journal Article · Tue Apr 01 04:00:00 EST 1986 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(86)90502-6· OSTI ID:5584925

Ludlow, J W; Guikema, J A; Consigli, R A

5-(4-Dimethylaminobenzylidene)rhodanine, a silver-specific dye, was used in a colorimetric assay to quantitate the autoradiographic deposition of silver onto X-ray film after exposure to sodium dodecyl sulfate-polyacrylamide gels of radiolabeled biological material. Silver grains were eluted from autoradiograms with 5 N potassium hydroxide, dissolved in nitric acid, and neutralized with 1 M Trizma Base. The concentration of silver was measured spectrophotometrically owing to the chelation properties of the dye. After corrections for background exposure were made, the silver contents of excised bands were then determined by comparison to a standard curve generated with silver nitrate. We have used this silver assay to quantitate the relative amount of each polypeptide band comprising the polyomavirus structural protein VP2 doublet. The method reported here has proven useful when densitometry is inconvenient (i.e., short distance between bands, irregular shape of bands, very faint bands) in addition to being inexpensive and simple to perform.

• Research Organization: Kansas State Univ., Manhattan
• OSTI ID: 5584925
• Journal Information: Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 154:1; ISSN ANBCA
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEAR CHEMISTRY
400103 -- Radiometric & Radiochemical Procedures-- (-1987)
400104* -- Spectral Procedures-- (-1987)
400702 -- Radiochemistry & Nuclear Chemistry-- Properties of Radioactive Materials
ALKALI METAL COMPOUNDS
AUTORADIOGRAPHY
BIOLOGICAL MATERIALS
CHELATES
CHEMICAL ANALYSIS
COLLOIDS
COMPLEXES
DEPOSITION
DISPERSIONS
DYES
ELECTROMAGNETIC RADIATION
ELEMENTS
ELUTRIATION
GELS
HYDROGEN COMPOUNDS
HYDROXIDES
IMAGE PROCESSING
INORGANIC ACIDS
IONIZING RADIATIONS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MATERIALS
METALS
MICROORGANISMS
NITRATES
NITRIC ACID
NITROGEN COMPOUNDS
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PARASITES
PEPTIDES
POLIO VIRUS
POLYMERS
POLYPEPTIDES
POTASSIUM COMPOUNDS
POTASSIUM HYDROXIDES
PROCESSING
PROTEINS
QUANTITATIVE CHEMICAL ANALYSIS
RADIATIONS
SEPARATION PROCESSES
SILVER
SILVER COMPOUNDS
SILVER NITRATES
SPECIFICITY
SPECTROPHOTOMETRY
TRACER TECHNIQUES
TRANSITION ELEMENT COMPOUNDS
TRANSITION ELEMENTS
VIRUSES
X RADIATION