Blotting description

Haselbeck, A. and Hosel, W. (1990) Description and application of an immunological detection system for analyzing glycoproteins on blots. Glycoconjugate J. 7, 63-74. 

The mRNA coding for 5-HT6 receptor has been localized in the rat brain by Northern blot, PCR, and in situ hybridization (206,207,213,214) (see also Fig. 10) and the protein by immunohistochemistry (213). The first and more detailed description on the localization of mRNA coding for 5-HT6 receptor was published by Ward and co-workers (215), reporting that the main rat brain regions where this receptor is expressed is the pyramidal layer of the olfactory tubercle, islands of Calleja, nucleus accumbens, striatum, hippocampus, and piriform cortex. At moderate levels, it is expressed in other cortical areas, the olfactory bulb, some nuclei of the hypothalamus and amygdale, the habenula, and the cerebellum. No mRNA expression is found in the raphe nucleus. These results were confirmed later (204). 

Before 1800, electricity meant static electricity, generated by friction. It could be stored in jar-like condensers, and a number of these condensers could be discharged simultaneously, like an artillery battery, producing a very hefty shock—up to half a million volts. The sparks from such discharges could ignite gas mixtures and decompose relatively small samples of some substances. Then in 1800, Volta published a description of a new piece of apparatus, the electric pile. It was called a pile because it consisted literally of a pile of alternating disks of metals and blotting paper moistened with a salt solution. It was... 

Specificity - A description of how the producer determined that the antibody binds only the listed antigen. Most of the time this is a western blot (with a blot shown), but it can be immunocytochemistry (with an image shown). Sometimes data are included about binding to other related proteins or to posttranslationally modified (e.g., phosphorylated) proteins. Some vendors who use a peptide for making an antibody will also sell the peptide for an absorption control. More information about specificity is discussed in the Chapter 9, Controls. Frequently, there are no data given for the specificity. 

The technique of Western blotting is widely used and a lot of investigations have been undertaken in order to quantify protein recovery on the collecting membrane [143, 144, 145, 146, 147]. One of the most common difficulties related to the description of the DPD transfer process is the estimation of the yield of proteins transferred from the gel onto the collecting PVDF membrane. One of the solutions was to use radiolabeUed proteins [193]. 

The dot-blot assay is a versatile tool its different modifications enable one to cope with almost every potentially interfering substance. In the following description the steps for all modifications are included. 

PSQ, 0.1 j,m pore size, Millipore, USA) and enclosed in a Western Blot cassette. After electro-transfer, the protein on the membrane is stained with a 0.1% Coomassie Brilliant Blue solution. The membrane is washed several times to remove residues from the SDS gel electrophoresis. The completeness of the transfer can be checked by staining the gel in the Coomassie Brilliant Blue solution. A detailed description of this procedure can be found in [369, 370). 

When this instrument was introduced to the market in about 1988 a description of the functioning of the apparatus was reported (62). The Wallac Ambis 40(X) directly detects P-particles from a wide variety of isotopes and is suitable for gels, blots, TLC plates, and any sample type of the dimension 20 x 20 cm. It is reported in the company literature that this instrument can be as much as 100 times faster than x-ray film. 

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