![]() These subclasses still contain the heavy (gamma) portion of the antibody, so IRDye subclass-specific antibodies still recognize them. In mice, 95% of light chains are kappa, and 5% are lambda. There are two types of light chains, kappa and lambda. Blots were detected using IRDye subclass-specific antibodies or IRDye whole IgG antibodies.Īntibody subclasses may also be designated by their light chains. Each lane was loaded with 50 ng of antibody. Western blot detection of various purified subclasses. Figure 96 demonstrates the differences in detection between IRDye antibodies. Other IRDye Secondary Antibodies are whole IgG (H + L) and react with the heavy (gamma) and light (kappa or lambda) chains of the primary antibody. Because of this, IRDye Goat anti-Mouse IgG 1, IRDye Goat anti-Mouse IgG 2a, and IRDye Goat anti-Mouse IgG 2b allow for two-color detection using primary antibodies derived from mice. For example, IRDye Goat anti-Mouse IgG 1 recognizes mouse gamma 1 it will not recognize mouse gamma 2a, 2b, 2c, or gamma 3. So, antibodies directed against a particular subclass will not recognize antibodies directed against other subclasses. IRDye subclass-specific antibodies only react with the heavy chain of a primary antibody. In mice, there are five unique subclasses of IgG:Įach subclass is based on small differences in amino acid sequences in the constant region of the heavy (gamma) chains. This guide will help you understand how to use IRDye® subclass-specific antibodies in your Western blot protocol. Using IRDye® Subclass-Specific Antibodies for Western Blot Detection Introduction
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