Flow cytometry was used to measure the binding of anti-α5β1 antibodies to HFF cells under conditions promoting low (in the presence of 1 mM each Ca²⁺ and Mg²⁺) and high affinity (in the presence of Mn²⁺ with or without the addition of 50K ligand). The non-function perturbing antibodies K20 (anti-β1) and mAb11 (anti-α5) were used to estimate total expression of each subunit. The results are expressed as mean fluorescence intensity (MFI) with background binding levels of normal rat or mouse IgG subtracted, and are representative of three separate experiments.

Neutral antibody.