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Isotype Controls
Isotype controls are non‑immune antibodies matched to the primary antibody’s host species, isotype, and conjugation, but lacking specificity for the target antigen. They are used as critical negative controls to quantify nonspecific binding (including Fc‑receptor–mediated binding), which is particularly important in FcR‑rich myeloid populations.
Quick Search: Select by Host Species and Subclass
Critical Selection Parameters
Parameter | Technical Rationale |
|---|---|
Ig Subclass | Different subclasses (e.g., IgG1 vs IgG2a) have distinct Fcγ‑receptor affinities and background profiles; best practice is to use an isotype control with the exact same subclass. |
Concentration | Nonspecific binding is governed by mass‑action; the control antibody should be matched to the primary by µg/mL concentration, not just by dilution factor. |
Fluorochrome | The fluorochrome and F/P ratio affect charge and nonspecific "stickiness"; the isotype control should use the same dye and a similar F/P ratio. |
Host Species | Matching host species ensures the secondary antibody recognizes control and primary identically; an exact species match is recommended. |
