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Blotting Western

The secondary antibody recognizes the primary antibody and binds at locations on the blot where the primary antibody is bound as well. The secondary antibody is furthermore conjugated with an enzyme or marked with a nucleotide, thus allowing detection (Fig. B.4). [Pg.29]

When using Alkaline Phosphatase based secondary antibody  [Pg.31]


Chemiluminescence and bioluminescence are also used in immunoassays to detect conventional enzyme labels (eg, alkaline phosphatase, P-galactosidase, glucose oxidase, glucose 6-phosphate dehydrogenase, horseradish peroxidase, microperoxidase, xanthine oxidase). The enhanced chemiluminescence assay for horseradish peroxidase (luminol-peroxide-4-iodophenol detection reagent) and various chemiluminescence adamantyl 1,2-dioxetane aryl phosphate substrates, eg, (11) and (15) for alkaline phosphatase labels are in routine use in immunoassay analyzers and in Western blotting kits (261—266). [Pg.275]

The technique of immun obi otting, often referred to as western blotting, is frequendy used for a variety of appHcations where protein concentrations... [Pg.184]

The protein was purified by a dialysis procedure, denatured and analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Western blotting indicated that the protein of interest consisted of two components, one of which increased in concentration as the purification proceeded. The authors initially suggested that this could be due to the presence of a number of species produced by modification of the amino acid side-chains, for example, by glyco-sylation, or by modification of the C- or N- terminus. [Pg.198]

Western blotting A means of transferring protein bands from an electrophoresis gel onto a fixing medium for further analysis. [Pg.312]

The venom of the Australian jack jumper ant M. pilosula has only been analyzed for its allergen composition in recent years. Besides three low molecular weight allergens, Myr pi, Myr p2 and Myr p3 (table 1), six higher molecular weight allergens of 22.8-89.9 kDa have been identified by Western blot [19]. Myr pi, a 25.6- and a 89.9-kDa protein could be major allergens. [Pg.147]

Western blot A method for transferring protein to a nitrocellulose filter, on which the protein can be detected by a suitable probe (eg, an antibody). [Pg.414]

A. Western blot analysis of membrane fractions from P. vulgaris probed with a PGIP-specific antibody. 1) PGIP purified from P. vulgaris] 2) and 3) membranes 4) supernatant of the membrane preparation. [Pg.200]

Further characterization by SDS-PAGE and Western blot analysis of the culture filtrate of a 6 day-old culture of lindemuthiaimm showed that only one endoPG of the same molecular weight (42 kDa) as the pure endoPG (Hugouvieux et al., 1995) was induced on pectin. [Pg.372]

Data represent the mean from 4- 6 tubers per line (Ig tissue per trial 3 replicons). Western blot analyses were positive. [Pg.391]

Figure 5, Western blot analyses. The blot was probed using antibodies raised against PL3. The PL3 protein bands corresponded to a molecular mass of 42 kDa. Figure 5, Western blot analyses. The blot was probed using antibodies raised against PL3. The PL3 protein bands corresponded to a molecular mass of 42 kDa.
Products were analysed by SDS-PAGE followed by Western blotting (Burnette 1981). RGase antibody production is described by van der Veen et al. 1991 This method can be used to determine the presence of RGase in different enzyme-preparations and culture-filtrates. [Pg.488]

Western blotting of above mentioned recombinant strains shows a clear increase of rhamnogalacturonase production compared to the wild strain. [Pg.491]

WN. Burnette (1981) "Western Blotting" Electrophoretic transfer of proteins from Sodium Dodecyl Polyacrylamide gels to unmodified nitrocellulose and radiographic... [Pg.494]

A. nidulans with selected phages of classes A, B, C, D and E resulted in increased polygalacturonase activity in comparison with the wild type strain. The culture media of pga transformants were further analysed by Western blotting with polyclonal antibodies raised against PGI and in all samples examined cross-reactive bands with molecular masses similar to that found for PGI or PGII were detected. [Pg.826]

The PemB cellular localisation was determined both in E. chrysanthenu and in an E. coli recombinant strain by Western blot of the cell fractions with a PemB-antiserum. No PemB was detected in the culture supernatant and only trace amounts were found in the soluble cell fractions - periplasm and cytoplasm (Figure 2). PemB was found mostly in the total membrane fraction from which it could be completely extracted by Triton X-100/Mg2+ and partially extracted by Sarkosyl (Figure 2). This behaviour is typical of inner membrane proteins, but since some exceptions have been noticed it does not positively indicate the PemB localisation (15). We performed cell membrane fractionation in sucrose density gradient centrifugation both by sedimentation and flotation, using several markers of inner and outer membrane vesicles. PemB was found in the outer membrane vesicles (data not shown). [Pg.839]

Specificity of the antisera was assessed by Western blotting. Electrophoretically separated proteins from culture filtrates were transferred to 0.45 fim nitrocellulose membranes. After transfer of proteins, membranes were... [Pg.883]

Both methods have been validated according to the Japanese validation protocol (Sakai et al., 2008), and both primers are commercially available. All the Western blotting and PCR kits are shown in Table 4.11. [Pg.156]

TABLE 4.11 Commercial Western blot and PCR kits for specific allergenic ingredients... [Pg.158]

Specific allergenic ingredient Western blot or PCR kits Target protein or gene Relative molecular weight (Da) or PCR product length (bp)... [Pg.158]

Egg Morinaga FASPEK Egg Western Blot Kit (ovalbumin) Morinaga Institute of Biological Sciences Co. Ovalbumin 50,000... [Pg.158]


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Antibodies Western blot

Antibodies Western blotting

Assay protein, Western blotting

Blots

Blots Blotting

Blotting

Chemiluminescence, enhanced Western blot

Enhanced chemiluminescence Western blotting

Experiment 18 Western Blot to Identify an Antigen

Far-Western blotting

Fusion western blot

General Carbohydrate Detection on Western Blots

Immunochemical techniques Western blotting

In Western blotting

North Western blotting

Peroxisome Western blot

Proteomics Western blots

Qualitative analysis Western blotting

Serum Glycoproteins by SDS-PAGE and Western Blotting

The Western Blot

Western

Western blot

Western blot

Western blot analysis

Western blot antibodies specificity determination

Western blot assay

Western blot blotting protocol

Western blot experiments

Western blot fractions

Western blot immunodetection

Western blot method

Western blot techniques

Western blot transfer procedure

Western blots hepatocytes

Western blots, ginsenosides

Western blotting analysis

Western blotting materials

Western blotting method

Western blotting primary antibodies

Western blotting technique

Western blotting technique sections

Western blotting/enzyme-linked

Western blotting/enzyme-linked ELISAs)

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