Mouse Anti-Alprenolol Recombinant Antibody (clone 37A4) (CAT#: FAMAB-0011CQ)

Figure 1 Serum anti-peptide response in rabbits immunized with HV3-Tg (∆ ,▲) to HV3-BSA conjugate (A) and to 37A4 Ig (B).

Preimmune serum (●)

Cazaubon, S., Couraud, P. O., Hoebeke, J., Nahmias, C., Emorine, L., Gras-Masse, H., & Strosberg, A. D. (1988). Anti-HV3 peptide antibodies as probes for conformational changes in immunoglobulins. Journal of immunological methods, 114(1-2), 13-20.

Figure 2 Inhibition of anti-HV3 binding to HV3-BSA conjugate (A) and to 37A4 Ig (B).

Antiserum was diluted at 1 : 50000 in PLT containing 0.5% BSA (A) and at 1 : 10000 in PLT (B). Inhibitors were HV3 (∆), 3qA4 (▲), SDS-denatured 37A4 (◊), SDS-denatured V66 (♦).

Cazaubon, S., Couraud, P. O., Hoebeke, J., Nahmias, C., Emorine, L., Gras-Masse, H., & Strosberg, A. D. (1988). Anti-HV3 peptide antibodies as probes for conformational changes in immunoglobulins. Journal of immunological methods, 114(1-2), 13-20.

Figure 3 Binding inhibition of anti-HV3 serum diluted at 1 : 10000 in PLT (b) to non-reduced (A) and reduced (B) 37A4 Ig.

Preimmune serum (a), Inhibitors were HV3 (c), 37A4 (d), V66 (e), SDS-denatured 37A4 (f), SDS-denatured V66 (g) each at 40μg/ml.

Cazaubon, S., Couraud, P. O., Hoebeke, J., Nahmias, C., Emorine, L., Gras-Masse, H., & Strosberg, A. D. (1988). Anti-HV3 peptide antibodies as probes for conformational changes in immunoglobulins. Journal of immunological methods, 114(1-2), 13-20.

Figure 4 Comparison between the percentage of inhibition of anti-HV3 serum to 37A4 by 0.1%, 0.3%, 0.9% SDS-denatured 37A4 at 40 μg/ml (□) (percentages calculated with respect to similarly treated V66) and the percentage of 0.1%, 0.3%, 0.9% SDS-denatured 37A4 binding with ³H-DHA (♦) (determined by the Farr technique).

Cazaubon, S., Couraud, P. O., Hoebeke, J., Nahmias, C., Emorine, L., Gras-Masse, H., & Strosberg, A. D. (1988). Anti-HV3 peptide antibodies as probes for conformational changes in immunoglobulins. Journal of immunological methods, 114(1-2), 13-20.

Figure 5 Binding isotherms at 27°C of ³H-l-dihydroalprenolol (○-○) and I-alprenolol (●-●) on the monoclonal antibody 37A4.

The first isotherm (○-○) was obtained by radioimmunoassay, the second isotherm (●-●) by fluorimetric titration. Both isotherms were normal&d for comparison purposes. The curves were computed by a non-linear regression (Wilkinson, 1961). The antibody combining site concn was 0.4 nM for the radioimmunoassay and 10 nM for the fluorescence titration. B/Bmax is the fractional saturation of antibody.

Hoebeke, J., Engelborghs, Y., Chamat, S., & Strosberg, A. D. (1987). The immune response towards beta-adrenergic ligands and their receptors—VII. Equilibrium and kinetic binding studies of l-alprenolol to a monoclonal anti-alprenolol antibody. Molecular immunology, 24(6), 621-629.

Figure 6 Association kinetics of ³H-l-dihydroalprenolol to 37A4 antibody.

(A) Binding as a function of time as tested at 27°C in a radioimmunoassay for four concns: 5 (○-○), 7.5 (●-●), 10 (□-□) and 12.5 nM (■-■). Equilibrium values are shown to the right of the plot. The antibody combining site concn was 0.4 nM. (B) Pseudo-first order plots of the association kinetics at 7°C (○-○), 17°C (●-●) and 27°C (□-□). kon was calculated from the slope and koff from the intercept on the y-axis.

Hoebeke, J., Engelborghs, Y., Chamat, S., & Strosberg, A. D. (1987). The immune response towards beta-adrenergic ligands and their receptors—VII. Equilibrium and kinetic binding studies of l-alprenolol to a monoclonal anti-alprenolol antibody. Molecular immunology, 24(6), 621-629.

Figure 7 Association kinetics of t-alprenolol to 37A4 antibody.

(A) Association curve obtained at 27°C for a concn of 7.5 μM antibody combining site and 120 μM alprenolol. The time scale was 5 msec/unit before and 10 msec/unit after the arrow. The ordinate is the fluorescence in arbitrary units. The insert shows the distribution of the residuals between the experimental and the calculated values for a single pseudo first order reaction. (B) Pseudo first order plots of the association kinetics at 7°C (◊-◊) (scale on the right y-axis), 17°C (×-×), 27°C (○-○) and 37°C (□-□) (scales on the left y-axis). The curve at 7°C was analysed as indicated in the Results.

Hoebeke, J., Engelborghs, Y., Chamat, S., & Strosberg, A. D. (1987). The immune response towards beta-adrenergic ligands and their receptors—VII. Equilibrium and kinetic binding studies of l-alprenolol to a monoclonal anti-alprenolol antibody. Molecular immunology, 24(6), 621-629.