Recombinant Anti-CDPK1 VHH Single Domain Antibody (PNBL-035) (CAT#: PNBL-035)

Figure 1 1B7 recognizes a calcium-dependent conformation of TgCDPK1

IRDye800-labeled 1B7 was used to probe decreasing concentrations of total T. gondii lysate or recombinant kinase (Top).Samples were separately probed for parasite actin (ACT1; red) or the 6xHis-tag on recombinant proteins (HIS; green).

Ingram, J. R., Knockenhauer, K. E., Markus, B. M., Mandelbaum, J., Ramek, A., Shan, Y., ... & Lourido, S. (2015). Allosteric activation of apicomplexan calcium-dependent protein kinases. Proceedings of the National Academy of Sciences, 112(36), E4975-E4984.

Figure 2 1B7 recognizes a calcium-dependent conformation of TgCDPK1

Incubation of parasite lysates with immobilized 1B7 depletes TgCDPK1/TgCDPK3 in the presence of CaCl2 but not in EGTA. The immunoprecipitated material could be eluted from the beads by subsequent incubation with EGTA. Blot was probed with 1B7 (green), and ACT1 (red) is included as a control.

Ingram, J. R., Knockenhauer, K. E., Markus, B. M., Mandelbaum, J., Ramek, A., Shan, Y., ... & Lourido, S. (2015). Allosteric activation of apicomplexan calcium-dependent protein kinases. Proceedings of the National Academy of Sciences, 112(36), E4975-E4984.

Figure 3 1B7 recognizes a calcium-dependent conformation of TgCDPK1

IP of recombinant kinases by 1B7 in the presence of 1 mM CaCl2 or EGTA. The domain structure of each kinase, indicating the KD and each of the EF hands (I–IV), is depicted for wild-type (WT), D368A/D415A (N-lobe mutant), and D451A/D485A (C-lobe mutant).

Ingram, J. R., Knockenhauer, K. E., Markus, B. M., Mandelbaum, J., Ramek, A., Shan, Y., ... & Lourido, S. (2015). Allosteric activation of apicomplexan calcium-dependent protein kinases. Proceedings of the National Academy of Sciences, 112(36), E4975-E4984.

Figure 4 1B7 recognizes a calcium-dependent conformation of TgCDPK1

Calcium dependency of TgCDPK1 activation or 1B7 binding. Values are normalized to maximal activity or binding.

Ingram, J. R., Knockenhauer, K. E., Markus, B. M., Mandelbaum, J., Ramek, A., Shan, Y., ... & Lourido, S. (2015). Allosteric activation of apicomplexan calcium-dependent protein kinases. Proceedings of the National Academy of Sciences, 112(36), E4975-E4984.

Figure 5 1B7 inhibits TgCDPK1 and related kinases.

In vitro kinase assay with increasing concentrations of 1B7.

Ingram, J. R., Knockenhauer, K. E., Markus, B. M., Mandelbaum, J., Ramek, A., Shan, Y., ... & Lourido, S. (2015). Allosteric activation of apicomplexan calcium-dependent protein kinases. Proceedings of the National Academy of Sciences, 112(36), E4975-E4984.