FabuLight - Fab Labeling Example Protocol
FabuLight™ Example Protocol - Fab Labeling Primary Antibodies
Include a blocking step if the tissue displays immunoglobulins that could be recognized by the FabuLight, for instance mouse tissue that is to be labeled with a mouse specific FabuLight. Otherwise start with step 2.
Incubate with unconjugated Fab anti-Fc antibody, in this example Fab Goat Anti-Mouse IgG1, Fcγ fragment specific. Fab Goat Anti-Mouse IgG (H+L) could also be used for this step. Wash.
2. Create Fab-primary antibody complexes. In this example Mouse Anti-Antigen X is complexed with Alexa Fluor® 488-conjugated Fab Goat Anti-Mouse IgG1, Fcγ fragment specific (Creating
Alexa Fluor® 488-labeled Anti-X Complex); and Mouse Anti-Antigen Y is complexed with Rhodamine Red-X-conjugated Fab Goat Anti-Mouse IgG1, Fcγ fragment specific (Creating Rhodamine Red-X-labeled Anti-Y Complex).
3. Incubate the sample with
Alexa Fluor® 488-labeled Anti-X Complex. (X being the less abundant target) Wash.
4. Incubate the sample with
Rhodamine Red-X-labeled Anti-Y Complex. (Y being the more abundant target) Wash.
Follow the steps above for an example protocol using Fab-labeled primaries for labeling two antigens on tissue.
Browse FabuLight - Affinity-Purified Fab Anti-Fc Fragment Specific Antibodies
Download White Paper - FabuLight Fab labeling primary antibodies with fluorophore-conjugated Fab anti-Fc