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BTighteners are used to obtain bright deposits direcdy from the bath (117). The additives currentiy used fall into two classes, which have vatiously been labeled primary and secondary, first class and second class, and carrier and brightener. The last is more commonly used in plating plants. [Pg.162]

In some cases, the preparation of a fluorescently labeled antibody is not even necessary. Particularly, if indirect methods are used to detect antibody binding to antigen, then preparing a fluorescently labeled primary antibody is not needed. Instead, the selection from a commercial source of a labeled secondary antibody having specificity for the species and class of primary antibody to be used is all that is required. However, if the primary antibody needs to be labeled and it is not manufactured commercially, then a custom labeling procedure will have to be done. [Pg.819]

Primary antibodies blot excess blocking solution from sections and incubate for 60 min at room temperature or overnight at +4°C with a mixture of correspondingly diluted unlabeled primary antibodies raised in two different host species (e.g., mouse and rabbit). When using fluorophore-labeled primary antibodies as in direct immunostaining method (one antibody layer), you may skip step (6) with secondary antibodies for indirect immunostaining method (two antibodies layers). Wash sections in PBS for 3 x 3 min. [Pg.71]

Another approach to this persistent problem relies on haptenylation of primary antibodies. Hapten (e.g., biotin, digoxigenin or any fluorophore) can be covalently bound to the antibody via A-hydroxysuccinimide esters (NHS-ES) (see Sect. 2.1), or conjugated employing monovalent IgG Fc-specific Fab fragments (see Sect. 2.2). Haptenylated primary antibodies can be subsequently visualized with the use of secondary antibodies recognizing the corresponding hapten (Fig. 8.5). Fluorophore-labeled primary antibodies can be directly visualized in a fluorescent microscope. [Pg.74]

Fluorophore absorbs ultraviolet light (or violet, blue or green) and emits light of longer wavelength. Fluorophores are used in immunohistochemistry for labeling primary or secondary antibodies in direct and indirect immunostain-ing methods, respectively. They can be visualized in fluorescence microscopy using special filter sets. [Pg.145]

Using an enzyme-labeled primary antibody, a very quick direct assay is obtained. This assay may not be as sensitive as others and may involve more... [Pg.184]

As shown in Fig. 21, in a direct ELISA the unlabeled antigen (a range of standard antigen concentrations or unknown samples) is attached to the solid phase. Enzyme-conjugated (labeled) primary antibody is then added. After incubation and washing of the plate... [Pg.395]

In labeling amino acids, peptides, and proteins, FITC can only label primary and secondary, but not tertiary, amino groups. Why (2 marks)... [Pg.397]

Incubation with a target protein binding protein (e.g., primary antibody) Wash three times with T-TBS or T-PBS for at least 5 min each. After washing, incubate the membrane with the (HRP-labeled) primary antibody solution for 2 h at room temperature or 37°C. [Pg.62]

Interestingly, hydroboration-amination as an isotope incorporation methodology has led to the synthesis of isomerically pure 13N- and 15N-labeled primary amines from labeled ammonia [23-25] (Scheme 8). These labeled amines are valuable intermediates in pharmacology. [Pg.43]

Simultaneous staining In a simultaneous double stain, the primary antibodies can be applied simultaneously. The advantage of this method is that it is less time-consuming because the reagents can be mixed together. However, the technique can only be used, if suitable primary antibodies are available. Two methods can be adopted A direct method with directly-labeled primary antibodies, or an indirect method based on unlabeled primary antibodies raised in different host species, or of different Ig isotype or IgG subclass (4). [Pg.105]

Multi-step technique (3) This is an indirect/direct method combining unlabeled primary antibodies with directly-conjugated antibodies. The method starts with staining the unlabeled antibody/antibodies with the appropriate detection system, but without performing the final enzymatic staining reaction. The tissue is blocked with normal serum from the host of the first primary antibody before the second, directly-labeled primary antibody is added. The staining ends with the two enzymatic reactions being performed sequentially. [Pg.105]

Immunohistochemical staining can be direct or indirect. Direct immunohis-tochemical staining methods utilize only a primary antibody, which may be conjugated to horseradish peroxidase, biotin, alkaline phosphatase, or other chromogens. In the case of biotin-labeled primary antibodies, avidin or strep-tavidin linked to peroxidase binds to the biotin allowing detection of reactivity of the test antibody with the tissue. Indirect immunohistochemical staining methods utilize secondary, tertiary, or even quaternary antibodies, any of which may be linked either to biotin or enzyme (e.g., peroxidase). [Pg.219]

The polyclonal antiserum, obtained by immunisation of rabbits as described in Materials and Methods was initially used for immunoprecipitation of metabolically labelled primary cortical neurons, providing a major band at 110 kDa corresponding to intact neuronal APP (Figure 2 A). The antibody was also tested by western analysis yielding... [Pg.344]

There is some confusion as to which electron acceptor should be called primary. Historically, in purple bacteria the quinone acceptor, Q, was so named. Later it was found that a BPh molecule accepts an electron before Q, and possibly even earlier acceptors, or charge transfer states, exist. Since the latter matter is still under debate (see Chapter 3), one might prudently keep the label primary for the quinone acceptor with the understanding that it is the first stable (on a time scale of ms) acceptor. [Pg.109]

Reduction of Aldehydes. Stereospecificially labeled primary alcohols are useful in biochemical and physical organic studies. Such compounds may be prepared by enzymatic reduction of a labeled aldehyde using yeast. However, isolation of the product is often tedious. Alpine-Borane greatly simplifies the process and provides compounds of high enantiomeric purity. It is the most efficient reagent available for reduction of aldehydes. The limiting... [Pg.478]

Figure 12.6 Schematics showing two common types of protein immunoassays. (Left) Direct assay exploiting interactions between target proteins and labeled primary antibodies. (Right) Indirect, sandwich assay relying on interactions, firstly between unlabeled primary antibodies and target proteins, and secondly between target proteins and labeled secondary antibodies. Figure 12.6 Schematics showing two common types of protein immunoassays. (Left) Direct assay exploiting interactions between target proteins and labeled primary antibodies. (Right) Indirect, sandwich assay relying on interactions, firstly between unlabeled primary antibodies and target proteins, and secondly between target proteins and labeled secondary antibodies.
When terpenic 1-deuterioaldehydes are subjected to the reduction with (R)- or (5)-BINAL-H, optically active, deuterium-labeled primary alcohols are produced (Scheme 4). ... [Pg.163]


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Labels direct primary

Labels indirect primary

Labels nonradioactive primary

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