Far-Eastern blotting
Far-Eastern blotting is a technique developed in 1994 by Taki and colleagues[1] at the Tokyo Medical and Dental University, Japan for the analysis of lipids separated by high-performance thin layer chromatography (HPTLC). The lipids are transferred from the HPTLC plate to a PVDF membrane for further analysis, for example by enzymatic or ligand binding assays[1] or mass spectrometry.[2]
Cholesterol, glycerophospholipids and sphingolipids are major constituents of the cell membrane and in certain cases function as second messengers in cell proliferation, apoptosis and cell adhesion in inflammation and tumor metastasis. Far-eastern blotting was established as a method for transferring lipids from an HPTLC plate to a polyvinyledene difluoride (PVDF) membrane within a minute. Applications of this with other methods have been studied. Far-eastern blotting allows for the following techniques:
- Purification of glycosphingolipids and phospholipids.
- Structural analysis of lipids in conjunction with direct mass spectrometry.
- Binding study using various ligands such as antibodies, lectins, bacterium, viruses, and toxins, and
- Enzyme reaction on membranes.
Not only analysis of lipids but also metabolites of drugs and natural compounds from plants, and environmental hormones are possible by this method.
Etymology
The name is a dual reference to Eastern blotting and the geographical concept of the "Far East" (which includes Japan).
References
- 1 2 D. Ishikawa & T. Taki (1998), "Micro-scale analysis of lipids by far-eastern blot (TLC blot)", Nihon yukagaku kaishi, 47 (10): 963–970, doi:10.5650/jos1996.47.963
- ↑ Hamasaki H, Aoyagi M, Kasama T, Handa S, Hirakawa K, Taki T (January 1999). "GT1b in human metastatic brain tumors: GT1b as a brain metastasis-associated ganglioside". Biochim. Biophys. Acta. 1437 (1): 93–9. doi:10.1016/S1388-1981(98)00003-1. PMID 9931455.