Western blotting/enzyme-linked

HIV diagnosis is made either by a positive HIV enzyme-linked immunosorbent assay (ELISA) or rapid test (these tests may be positive as soon as 3 to 6 weeks after infection) and then confirmed by a positive confirmatory test, usually the HIV Western blot (Table 84-1). 

The use of independent methods, other than IFIC, for quantitative demonstration of proteins is particularly important. Both enzyme-linked immunosorbent assay (ELISA) and Western blot may be employed to confirm the amount of protein in a cell/tissue model, and in the protein-embedding bar code model under both comparable fresh and FFPE samples for accurate... 

Positive enzyme-linked immunosorbent assays are repeated in duplicate and if one or both tests are reactive, a confirmatory test is performed for final diagnosis. Western blot assay is the most commonly used confirmatory test, although an indirect immunofluorescence assay is available. 

PAMAM dendrimers or modified PAMAM dendrimers (with oxiamine or sulfhydryl surface functionalities) exclusive of one another. These antisera recognize den-drimers in ELISA (enzyme-linked immunosorbent assays), dot blots and Western blots. The immunogenicity of dendrimer-protein conjugates has implications for therapeutic use of dendrimers as vaccines and we anticipate that antidendrimer antibodies will have applications in patterning and assembling nanostructures containing dendrimers. 

Immune detection is a key utility of antibodies in biotechnology [3, 5]. Antiden-drimer sera efficiently detect dendrimers in multiple assay formats, including enzyme-linked immunosorbent assays (ELISA), and in Western and dot blots [3, 5], ELISA assays are commonly used to quantitate proteins, and a quantitative ELISA could be developed for dendrimers using our sera, though doing so would require development of dendrimer standards of known concentration that could be used for calibration. 

Testing for HIV exposure by ELISAs (enzyme-linked immunosorbent assays) and western blots. 

The products of gene expression (mRNA and proteins) can be measured by techniques such as the following. Northern blots are very similar to Southern blots except that the original sample contains a mixture of mRNA molecules that are separated by electrophoresis, then hybridized to a radioactive probe. Microarrays are used to determine the differing patterns of gene expression in two different types of cells—for example, normal and cancer cells. Enzyme-linked immunosorbent assays (ELISAs) and western blots (immunoblots) are used to detect specific proteins. 

This can be done by the Ouchterlony diffusion technique (see Fig. 2 and ref. 6). by enzyme-linked immunosorbent assay (see Chapter 15), or by Western blot, either using the purified protein or a more complex mixture of proteins containing the antigen of interest separated on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel (see Chapter 20). 

Three main techniques have been used for detecting HER-2 oncoprotein overexpression Western blot analysis, enzyme-linked immunosorption assay (ELISA), and immunohistochemistry. Western blot analysis is limited to basic research rather than routine clinical... 

It is highly recommended that microarray findings be verified using other experimental approaches. We usually confirm our results by at least one of the alternative immunoassays, such as enzyme-linked immunosorbent assay (ELISA), dot blot, Western blot, flow cytometry, or immunohistology (16,18). 

A standard double-sandwich enzyme-linked immunosorbent assay (ELISA) or Western blot analysis is used. As the concentration of factor is very low in normal plasma (approximately 0.1 mg/1 for factor VIII), it is necessary to subject plasma samples to cryoprecipitation in order to concentrate the sample, prior to Western blot analysis. The cryoprecipitation protocol described by Bi et al. is as follows (Bi et al., 1996 Sarkar et al., 2000 Mah et al., 2003). Plasma samples are collected as described above for the Coatest assay. Plasma is then frozen at -80 °C overnight. Frozen samples are then subject to centrifugation at 7000 x g for 20 min at 4°C. The precipitate is washed with... 

For cell surface antigens, the most frequently used assays are immunofluorescence or immunoenzymatic, with their numerous variations. For soluble protein or peptide antigens, the most common assays are immunoenzymatic, enzyme-linked immunosorbent assays (ELISA) or Western blot tests. 

Cultured cells are treated with various doses of test substances in varying time periods. After the completion of treatment, they are harvested and lysed. The lysates are analyzed for p53 by Western blot and/or enzyme-linked immunosorbent assay analysis. An increase in cellular p53 following treatment is interpreted... 

To examine the purity of the protein that was eluted from a Sephacryl S-300 column, we subjected the protein to SDS polyacrylamide gel electrophoresis (SDS-PAGE) and then performed a Western blot (26) using avidin linked to phosphatase as a probe. The SDS-PAGE indicated that there were many polypeptides in this partly purified preparation (Fig. 6). Among these were two major and several minor biotin-containing bands (Fig. 7). This preparation may contain propionyl CoA carboxylase or a partly degraded form of ACCase that can use propionyl CoA as well as acetyl CoA as a substrate. Therefore, the enzyme data presented here are quali-... 

One of the most important requirements for successful protein purification is the availability of an accurate, rapid, and quantitative assay that is specific for the protein of interest. If the protein has no known enzymatic activity, the amount of the protein present after each purification step must be analyzed by some general method such as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting, or an enzyme-linked immunosorbent assay (ELISA). If... 

Enzyme-linked immunosorbent assay (ELISA) Section 4.3.3 Western blotting Section 4.3.4... 

Proteins can be detected and quantitated by highly specific antibodies monoclonal antibodies are especially useful because they are homogeneous. Enzyme-linked immunosorbent assays and Western blots of SDS-polyacrylamide gels are used extensively. Proteins can also be localized within cells by immunofluorescence microscopy and immunoelectron microscopy. 

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