MALDI-TOFMS Identification of ‘Odorant Binding Proteins’ (OBPs) Electroblotted onto Poly(vinylidene difluoride) Membranes
✍ Scribed by J. C. Blais; P. Nagnan-Le-Meillour; G. Bolbach; J. C. Tabet
- Book ID
- 102650103
- Publisher
- John Wiley and Sons
- Year
- 1996
- Tongue
- English
- Weight
- 584 KB
- Volume
- 10
- Category
- Article
- ISSN
- 0951-4198
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✦ Synopsis
Matrix-assisted laser desorptionlionization time-of-flight mass spectrometry (MALDI-TOFMS) analysis of proteins electroblotted onto membranes has been explored for identification of odorant-binding proteins (OBPs) extracted from M. brassicae antennae and separated by native polyacrylamide gel electrophoresis (PAGE). At 337 nm laser wavelength, the best results have been obtained with an Immobilon P membrane and a-cyano-4-hydroxycinnamic acid as matrix. The values of the molecular weights of the different OBP's present in the electrophoretic bands have been determined with an accuracy of 0.1 % to 0.3% using bovine insulin as an internal standard. These results are in good agreement with estimations obtained by SDS-PAGE and bring confirmation of previous hypotheses concerning the structure of these OBPs.
EXPERIMENTAL
Polyacrylamide gel electrophoresis separation and electroblotting of proteins
Proteins were extracted from 3-day old male and female M. brassicae. Protein separation by native-PAGE used the procedure previously described by Nagnan-Le Meillour et