Mixed-substrate (glycerol tributyrate and fibrin) zymography for simultaneous detection of lipolytic and proteolytic enzymes on a single gel = 단일 젤에서 두 가지 효소의 동시 감지를 위한 혼합기술 자이모그라피

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Title
Mixed-substrate (glycerol tributyrate and fibrin) zymography for simultaneous detection of lipolytic and proteolytic enzymes on a single gel = 단일 젤에서 두 가지 효소의 동시 감지를 위한 혼합기술 자이모그라피
Author(s)
Nack-Shick Choi; Jong Hyun Choi; B H Kim; Yunjon HanJoong Su KimSeung Goo LeeJae Jun Song
Bibliographic Citation
Electrophoresis, vol. 30, no. 12, pp. 2234-2237
Publication Year
2009
Abstract
A new zymography method for simultaneous detection of two different enzymatic activities (lipolytic and proteolytic) using a single SDS-containing or native-conformation gel and a mixed-substrate (glycerol tributyrate and fibrin) (MS)1 gel was developed. After routine electrophoresis, SDS in the gel was removed by treatment with Triton X-100. Gel proteins were electrotransferred to the MS gel. To visualize lipolytic activity, the MS gel was incubated at 371C (for 6 or 24 h) until clear bands against an opaque background were observed. To detect proteolytic activity, the same MS gel was stained with Coomassie brilliant blue. Using this method, we show that six lipolytic enzymes from Staphylococcus pasteuri NJ-1 and four proteolytic enzymes from two Bacillus strains, B. licheniformis DJ-2 and B. licheniformis NJ-5, isolated from soil, can be simultaneously detected.
Keyword
ElectrotransferLipaseMixed-substratesProteaseZymography
ISSN
0173-0835
Publisher
Wiley
DOI
http://dx.doi.org/10.1002/elps.200800727
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
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