CyDye Immunoblotting for Proteomics: Co-detection of specific immunoreactive and total protein profiles
نویسندگان
چکیده
The development of ECL-Plex CyDye-conjugated secondary antibodies allows the advancement of conventional Western blotting, opening up possibilities for highly sensitive and quantitative protein confirmation and identification. We report a novel proteomic method to simultaneously visualise the total protein profile as well as the specific immunodetection of an individual protein species by combining cyanine CyDye pre-labelled proteins and antibody immunoblotting. This technique proposes to revolutionise both 2-D immunoprobing and protein confirmation following MS analysis.
منابع مشابه
Isolation and identification of excretory-secretory and somatic antigens from the Oestrus ovis larvae by SDS-PAGE and immunoblotting
Oestrus ovis is an economically important parasite of small ruminants and a zoonotic parasite with many reports of ophthalmomyiasis in human from Iran and other countries. The aim of the peresent studywas the isolation and identification of excretory-secretory (ES) and somatic (S) antigens of O. ovis second and third stage larvae (L2, L3) collected from Arabi sheep breeds loca...
متن کاملO-5: Identification of Novel ImmunodominantEpididymal Sperm Proteins Using CombinatorialApproach
Background: Alteration in the protein signatures of functionally immature testicular spermatozoa occurs during their journey through the epididymis. This leads to acquisition of sperm domain specific functions essential for successful fertilization. Epididymal sperm proteins are preferred targets for immunocontraception as well as in elucidating the causes of infertility. The Background of the ...
متن کاملProteomic and immunoassay characterization of a new food allergen from hazelnut (corylus avellana)
Methods Otherwise healthy children with oral food challenge confirmed hazelnut allergy were prospectively evaluated. Crude protein extracts were obtained from 5 hazelnut varieties, including autochthon, Northern Italy and Oregon (USA) cultivars, with phosphate saline buffer, pH 7.2. The immunoreactive protein components were identified by SDS-PAGE electrophoresis and Western immunoblotting, usi...
متن کاملDetection and mass spectrometry identification of protein changes in low-abundance tissue using CyDye DIGE Fluor saturation dyes
Traditional two-dimensional electrophoresis (2-DE) is a wellestablished technique for protein analysis, but its use in identifying changes in protein abundance in tissue samples has been limited by system variability and low sensitivity. These limitations have largely precluded its use to identify protein differences in scarce materials such as those recovered from laser-capture microdissection...
متن کاملProteomic Analysis of Gene Expression in Basal Cell Carcinoma
Background: Basal Cell Carcinoma (BCC) is a type of non-melanoma skin cancer. Alteration in gene expression is the important event that happens in cancer cell. Detection of this event is possible by proteomics techniques. Methods: Normal and tumor tissues were taken from BCC patient. Total proteins were purified by standard methods, and proteins were separated by two-dimensional electrophoresis...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 6 شماره
صفحات -
تاریخ انتشار 2006