Mouse Anti-TSHR Recombinant Antibody (clone 4G11) (CAT#: FAMAB-0323CQ)

Figure 1 D2 and 4G11 monoclonal antibodies specifically interact with the TSH receptor on FRTL-6 and TSH receptor-transfected FRT cells.

Panel A, ability of D2 and 4G11 antibody preparations to bind to membrane preparations from FRTL-5 rat thyroid cells as a function of concentration. Binding was measured by an enzyme-linked immunosorbent assay assay. Binding is expressed by the increase in optical density effected by the detection antibody reactivity with substrate resulting in increased absorbance at 415 nm. Data represent the mean of triplicate assays from a single experiment and S.E. is indicated; a duplicate experiment yielded identical data using a different batch of cells. The control IgM in this experiment was the affinity purified mouse IgM from Zymed. The same results were obtained with two monoclonal IgM preparations: IgM, K (Chemicon), and the control IgM used previously. Statistically significant (p < 0.01) binding by comparison to FRT cells with or without normal mouse IgM. Panel B, effect of increasing concentrations of TSH on the binding of D2 and 4G11 to FRTL-5 thyroid membrane preparations. Statistically significant inhibition of binding (p < 0.01). Panel C, binding of D2 and 4G11 antibodies to membranes from FRT cells (FRT cells) which have no functional TSH receptor and no TSH receptor mRNA to membranes from T33 FRT cells (TSH receptor-transfected FRT cells) which contain transfected TSH receptor cDNA and regain TSH binding and TSH-stimulated adenylate cyclase activity, and to membranes from T33 cells treated with 10⁻⁷ M TSH, (TSH-receptor-truncted FRT cells + 10⁻⁷ M TSH). The data represent the mean value of quadruplicate results from a single experiment ± S.E. Two separate experiments using different batches of cells yielded similar results. Where relevant, statistically significant binding (p < 0.01) and inhibition of binding (p < 0.01) by TSH were observed.

Taub, R., Hsu, J. C., Garsky, V. M., Hill, B. L., Erlanger, B. F., & Kohn, L. D. (1992). Peptide sequences from the hypervariable regions of two monoclonal anti-idiotypic antibodies against the thyrotropin (TSH) receptor are similar to TSH and inhibit TSH-increased cAMP production in FRTL-5 thyroid cells. Journal of Biological Chemistry, 267(9), 5977-5984.

Figure 2 Ability of peptides to inhibit the binding of the D2 (A) or 4G11 (B) antibodies to membrane preparations from FRTL-5 rat thyroid cells as a function of peptide concentration.

Binding was measured by an enzyme-linked immunosorbent assay. Binding is expressed by the increase in optical density effected by the detection antibody reactivity with substrate resulting in increased absorbance at 415 nm. The mean of three separate experiments and the S.E. is indicated. In all experiments, all of the unrelated peptides (in A: P3, P5, P7; in B: PI, P4, P6) were used as controls, although the data for only one unrelated peptide is shown. The x-axis represents a logarithmic scale of peptide concentration.

Taub, R., Hsu, J. C., Garsky, V. M., Hill, B. L., Erlanger, B. F., & Kohn, L. D. (1992). Peptide sequences from the hypervariable regions of two monoclonal anti-idiotypic antibodies against the thyrotropin (TSH) receptor are similar to TSH and inhibit TSH-increased cAMP production in FRTL-5 thyroid cells. Journal of Biological Chemistry, 267(9), 5977-5984.

Figure 3 Ability of peptides related to D2 (A) or 4G11 (B) to inhibit the ability of 5 X 10⁻¹⁰ M TSH to increase cAMP levels in rat FRTL-5 thyroid cells.

Taub, R., Hsu, J. C., Garsky, V. M., Hill, B. L., Erlanger, B. F., & Kohn, L. D. (1992). Peptide sequences from the hypervariable regions of two monoclonal anti-idiotypic antibodies against the thyrotropin (TSH) receptor are similar to TSH and inhibit TSH-increased cAMP production in FRTL-5 thyroid cells. Journal of Biological Chemistry, 267(9), 5977-5984.