Heavy chains, antisera

In monoclonal gammopathies, the IFE patterns usually yield a distinct, sharply defined precipitin band with one heavy-chain and one light-chain antiserum. These bands match the location of the particular immunoglobulin in the reference pattern (Figure 20-10). A second, fainter band of free light chains may also be present. 

Fig. 2. TrkA associates with p75 receptor in 293T cells. 293T cells ( 1 X 10 ) were co-transfected with p75 and either TrkA or empty vector. 36 h after transfection, the cells were harvested into TNE buffer [10 mM Tris (pH 8.0), 150 mM NaCl, 1 mM EDTA, and 1% NP40], The lysates were immunoprecipitated with anti-pan Trk antibody (Cl4, Santa Cruz) which recognizes the C-terminus of Trk and subsequently subjected to Western blot analysis with an anti-p75 antiserum (9992), or anti-pan Trk antiserum. The amount of p75 protein for each transfection is assayed in the bottom panel. The protein visualized in both lanes of the top panel reflects reactivity of the heavy chain of IgG.

These deseriptions deal with the processing of the donor serum after production and before conjugation. The description of the immimogen is also sometimes critical. Using the last example, the cow IgG may have itself been affinity purified before injection of the donkey, or specific parts of the Ig purified to raise a subclass specific antiserum, for example, heavy chain IgG may have been... 

Feizi et al. (1977) have described a heavy-chain variable region marker (VnMar) that was detected using an antiserum raised against human IgM cold agglutinins. Evidence was presented that the relevant determinant was associable with cross-reactive idiotypes characteristic of the cold agglutinins. It was of particular interest that this antigen is found in both normal pooled IgG and some unselected myeloma paraproteins. 

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