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A new method for the detection of proteolytic activity in Pseudomonas lundensis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Electrophoresis. 1995 Jan; 16(1):43-5.E

Abstract

A new method for the visualization of proteolytic activity in cell culture supernatant from Pseudomonas lundensis after sodium dodecyl sulfate (SDS)--gel electrophoresis is described. Following conventional electrophoresis, the gel is washed in a methanol-containing buffer to facilitate partial removal of SDS. After incubation with 0.5% casein the gel is stained for protein with Coomassie Brilliant Blue R-250. Bands with proteolytic activity appear as clear areas in the gel against a blue-stained background. Molecular weight standards electrophoresed in the same gel stain more intensely than the background and allow determination of the molecular weights of the proteolytic components. The sensitivity of post-electrophoretic reactivation in SDS-gels was determined using trypsin as standard. A slight modification of the technique allowed detection of proteolytic activity in nondenaturing and in isoelectric focusing gels.

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Publisher Full Text

Authors+Show Affiliations

Lundy FT
Food Studies Research Unit, University of Ulster at Jordanstown.
Magee AC
No affiliation info available
Blair IS
No affiliation info available
McDowell DA
No affiliation info available

MeSH

CaseinsCulture MediaElectrophoresis, Polyacrylamide GelEndopeptidasesEnzyme ActivationIsoelectric FocusingPseudomonasPseudomonas fluorescensSensitivity and SpecificityTrypsin

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

7737090

Citation

Lundy, F T., et al. "A New Method for the Detection of Proteolytic Activity in Pseudomonas Lundensis After Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis." Electrophoresis, vol. 16, no. 1, 1995, pp. 43-5.
Lundy FT, Magee AC, Blair IS, et al. A new method for the detection of proteolytic activity in Pseudomonas lundensis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Electrophoresis. 1995;16(1):43-5.
Lundy, F. T., Magee, A. C., Blair, I. S., & McDowell, D. A. (1995). A new method for the detection of proteolytic activity in Pseudomonas lundensis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Electrophoresis, 16(1), 43-5.
Lundy FT, et al. A New Method for the Detection of Proteolytic Activity in Pseudomonas Lundensis After Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis. Electrophoresis. 1995;16(1):43-5. PubMed PMID: 7737090.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A new method for the detection of proteolytic activity in Pseudomonas lundensis after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AU - Lundy,F T, AU - Magee,A C, AU - Blair,I S, AU - McDowell,D A, PY - 1995/1/1/pubmed PY - 1995/1/1/medline PY - 1995/1/1/entrez SP - 43 EP - 5 JF - Electrophoresis JO - Electrophoresis VL - 16 IS - 1 N2 - A new method for the visualization of proteolytic activity in cell culture supernatant from Pseudomonas lundensis after sodium dodecyl sulfate (SDS)--gel electrophoresis is described. Following conventional electrophoresis, the gel is washed in a methanol-containing buffer to facilitate partial removal of SDS. After incubation with 0.5% casein the gel is stained for protein with Coomassie Brilliant Blue R-250. Bands with proteolytic activity appear as clear areas in the gel against a blue-stained background. Molecular weight standards electrophoresed in the same gel stain more intensely than the background and allow determination of the molecular weights of the proteolytic components. The sensitivity of post-electrophoretic reactivation in SDS-gels was determined using trypsin as standard. A slight modification of the technique allowed detection of proteolytic activity in nondenaturing and in isoelectric focusing gels. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/7737090/A_new_method_for_the_detection_of_proteolytic_activity_in_Pseudomonas_lundensis_after_sodium_dodecyl_sulfate_polyacrylamide_gel_electrophoresis_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0173-0835&date=1995&volume=16&issue=1&spage=43 DB - PRIME DP - Unbound Medicine ER -