To minimize cross-reactivity, the donkey anti-mouse IgG whole antibodies have been cross-adsorbed against IgG from bovine, goat, chicken, guinea pig, hamster, horse, sheep, rabbit, rat, and human. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in less background staining and cross-reactivity. The secondary antibody solution is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Only the nonspecific-binding secondary antibodies are captured in the column, and the highly specific secondaries flow through. Further passages through additional columns result in highly cross-adsorbed preparations of secondary antibody. The benefits of these extra steps are apparent in multiplexing/multicolor-staining experiments where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there may be the presence of endogenous immunoglobulins.
Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining. Because staining protocols vary with application, the appropriate dilution of antibody should be determined empirically.
Specificity: This antibody binds to heavy chains on mouse IgG and light chains on all mouse immunoglobulins. This antibody does not bind non-immunoglobulin mouse serum proteins or IgG from bovine, chicken, goat, guinea pig, hamster, horse, human, rabbit, rat or sheep.
Obtain superior images with the new Invitrogen™ Alexa Fluor™ Plus secondary antibodies. Make your low abundant targets visible, minimize time spent optimizing, and make your precious samples count. Alexa Fluor™ Plus secondary antibodies provide brighter signal while having lower cross-reactivity compared to leading Alexa Fluor secondary antibodies, providing you with higher sensitivity and better signal-to-noise for your critical experiments. Alexa Fluor Plus secondary antibodies represent an advancement in fluorescent conjugate technology. Alexa Fluor Plus secondary antibodies are conjugated using new, proprietary dye chemistry and are pre-adsorbed to reduce background and cross-reactivity so you can generate stunning images. Alexa Fluor Plus secondary antibodies provide up to 4.2 fold higher signal-to-background in imaging applications and up to 5.8 fold higher signal-to-background in fluorescent western applications compared to Alexa Fluor secondary conjugates.
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