| Anti-IgG, Light Chain Specific for Western Blotting | ||||||||||||||||||||||||||||||||||||||||||||||||||
Anti-IgG, Light Chain Specific antibodies react with native primary antibodies used for detecting specific protein bands on Western blots. If diluted properly anti-light chain specific antibodies do not bind to the reduced and denatured IgG heavy chain band (50 kD) on blots (Figures A, C, and D). Therefore, by using anti-light chain specific antibodies, detection of antigens with molecular weights near 50 kD is not obscured by large amounts of reduced and denatured IgG heavy chains from primary antibodies used for immunoprecipitating (IP) primary antibodies (for example, see Figure B). |
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Figures A-D. Heavy (50 kD) and light (25 kD) chains of reduced and SDS-denatured mouse IgG(A-B), rat IgG (C), and rabbit IgG (D) were separated by SDS-PAGE (lanes with red numbers) and detected on Western blots using HRP-goat anti-mouse IgG, Light Chain specific (A), HRP-goat anti-mouse IgG (H+L)(B), HRP-Goat anti-rat IgG, Light Chain specific (C), and HRP-mouse anti-rabbit IgG, Light Chain specific (D). No heavy chain band was; detected even on lanes heavily overloaded with IgG when anti-IgG, Light Chain specific antibodies were used (A, C, and D) for detection. However, both heavy and light chain bands were detected with anti-IgG (H+L)(B). Lanes with blue numbers contained reduced and SDS-denatured goat IgG (A, B, and C) or mouse IgG (D), which served as background controls. |
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Although the antibodies react strongly with native IgG light chains, they do not react as strongly with the reduced and denatured light chains on blots. Therefore, they are not recommended for sensitive and quantitative detection of reduced and denatured light chains on Western blots. The antibodies have been thoroughly adsorbed to minimize cross-reactivity with immunoglobulins from many other species, which also may be present on blots. If the protein of interest has a reduced and denatured molecular weight near 25 kD, anti-IgG, Fc fragment specific antibodies may be used to detect native IgG primary antibodies without binding to the 25 kD band of reduced and denatured IgG light chains on Western blots. |
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* This antibody reacts primarily with kappa light chains. It is not suitable for detection of primary antibodies with lambda light chains. |
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