Immunofluorescence continued
      

Choice of Fluorophores. We offer secondary antibodies and avidin and streptavidin conjugated to traditional fluorochromes such as fluorescein, rhodamine, Texas Red®, AMCA, phycoerythrin – as well as the newer DyLight® dyes.

Species Cross-Reactivity Considerations. It is important to consider not only the species of the primary antibody but also the species of the tissue when choosing the optimal detection system for your application. If the species of the primary antibody and the species of the tissue are closely related (e.g. rat and mouse), the secondary antibody may bind to endogenous immunoglobulins in the tissue section leading to background. The following are options to minimize background staining in these instances: Use directly labeled primary antibodies. They can be labeled with either fluorescein (ProtOn™ Fluorescein Labeling Kit, PLK-1201) or with biotin (ProtOn™ Biotin Labeling Kit, PLK-1202). Biotinylated primary antibodies can be detected with a flurochrome-labeled avidin or streptavidin. Use a secondary antibody specifically adsorbed to remove cross-reacting antibodies of closely-related species (e.g. fluorescein conjugated anti-mouse IgG, rat adsorbed, FI-2001). Use the M.O.M.™ Immunodetection System for applications of mouse primary antibodies on mouse tissue. Multiple Antigen Labeling. Our affinity purified, highly adsorbed secondary antibodies and our avidin and streptavidin conjugates allow specific and crisp labeling of multiple antigens in the same section, as well as co-localization of antigens in the same cellular compartment of a section. When choosing detection systems for double antigen labeling, the species of each primary and secondary antibody, as well as the tissue species, must be considered. If the two primary antibodies are both made in mouse, directly label and detect the second mouse primary antibody, or use the M.O.M.™ Immunodetection System. The M.O.M™ system contains a proprietary Mouse Ig Blocking Reagent and a specially modified biotinylated anti-mouse secondary antibody. Together, these reagents minimize cross-reactivity of the detection system for endogenous mouse IgG. Used in a double-label application with two mouse primaries, this system will minimize cross-reactivity of the second detection system with the first, resulting in specific and discreet staining of the two antigens. For more information, request or download our free brochure on multiple antigen labeling, “Discovery Through Color”. VECTASHIELD® Mounting Media. VECTASHIELD® Mounting Media are unsurpassed in preventing photobleaching. These ready-to-use mounting media are stored at 4 ºC, available in non-hardening and hardening versions, and with and without nuclear counterstains.