Mouse Anti-PRLR Recombinant Antibody (clone A8) (CAT#: PABZ-179)

Figure 1 Inhibition of sAB binding by hGH or PRL using phage ELISA.

Both hGH (left panel) and PRL (right panel) inhibit the interaction between phage-displayed sABs and hPRL-R in a concentration dependent manner.

Rizk, S. S., Kouadio, J. L. K., Szymborska, A., Duguid, E. M., Mukherjee, S., Zheng, J., ... & Kossiakoff, A. A. (2015). Engineering synthetic antibody binders for allosteric inhibition of prolactin receptor signaling. Cell Communication and Signaling, 13(1), 1.

Figure 2 sAB inhibition of hPRL-R in T47D breast cancer cells.

a-b Fluorescence microscopy images of cells treated with 100 nM cy5-hPRL (a) or cy5-hGH (b) in the presence of each sAB. All panels represent two merged channels; blue: DAPI nuclear stain, red: cy5. c A Stat5 phospholyration luciferase reporter assay of T47D cells in the presence or absence of hPRL and hPRL-R sABs. A sAB against bacterial maltose binding protein (MBP) was used as a negative control. d Western blot detection of the time-dependent inhibition of hPRL-R signaling by hPRL-R sABs.

Rizk, S. S., Kouadio, J. L. K., Szymborska, A., Duguid, E. M., Mukherjee, S., Zheng, J., ... & Kossiakoff, A. A. (2015). Engineering synthetic antibody binders for allosteric inhibition of prolactin receptor signaling. Cell Communication and Signaling, 13(1), 1.

Figure 3 Photo-cross-linking of PapA8-DKP-insulin and control analogues to (A) the isolated ectodomain and (B) the lectin-purified holoreceptor.

Analysis of photoproducts by SDS-PAGE and Western blotting using NeutrAvidin (NAv) to detect the biotin tag on the insulin B chain (boxes a and c in each panel) or polyclonal antiserum (IRR-N) to detect N-terminal residues of the R-subunit (box b). (A) Photo-cross-linking via positions B25 (lanes 4 and 10), B16 (lane 8), and A8 (lane 14) analyzed following DTT reduction. Lanes 1-3, 5-7, 9, and 11-13 show control reaction mixtures in which either the ectodomain had been omitted (lanes 1, 2, 5, 6, 11, and 12) or samples had not been irradiated (lanes 1, 3, 5, 7, 9, 11, and 13). (b and c) Control blots to demonstrate that equal amounts of ectodomain (b, with DTT) and insulin derivatives (c, without DTT) were present in each reaction. (B) Photo-cross-linking to WGA-purified IR: (a) photo-cross-linking via positions B25 (lane 4), B16 (lane 8), A8 (lane 12), and B0 (lane 16) analyzed following reduction by DTT. Lanes 1-3, 5-7, 9-11, and 13-15 show control reaction mixtures in which either the IR had been omitted (lanes 1, 2, 5, 6, 9, 10, 13, and 14) or samples had not been irradiated (lanes 1, 3, 5, 7, 9, 11, 13, and 15). Boxes b and c are control blots as in panel A. Molecular masses are in each case shown at the right.

Wan, Z., Xu, B., Huang, K., Chu, Y. C., Li, B., Nakagawa, S. H., ... & Weiss, M. A. (2004). Enhancing the activity of insulin at the receptor interface: crystal structure and photo-cross-linking of A8 analogues. Biochemistry, 43(51), 16119-16133.