Enzymes

Complex of recombinant, full length human CDK4 (GenBank accession number NM_000075) with C-terminal His-tag and modifications (a.a. 43G-47G were replaced with EE, and T172A), and human Cyclin D1 (GenBank accession number NM_053056), a.a. 16-271, were co-expressed in a baculovirus-infected Sf9 cells expression system. CDK4 MW=34 kDa, Cyclin D1 MW=29 kDa.

Recombinant full-length human CDK5 (59 kDa) and human p35 (60 kDa), both with N-terminal GST-tags, were co-expressed in Sf9 cells using a baculovirus expression system. The CDK5 gene accession number is NM_004935 and p35 gene accession number is NM_003885.

Complex of recombinant human CDK6, full length with N-terminal His-tag, and Cyclin D1, full length with N-terminal GST-tag (GenBank Accession Nos. NM_001259, and NM_053056 respectively) co-expressed in Sf9 insect cells via a baculovirus expression system.

Complex of recombinant human CDK7, Cyclin H1 and MNAT1, full length, with N-terminal His-tags (CDK7 and MNAT1) and C-terminal His-tag (CyclinH1) , (GenBank Accession Nos. NM_001799, NM_001239, and NM_002431 respectively), co-expressed in Sf9 insect cells via a baculovirus expression system.

Human ULK1 (UNC-51-Like Kinase-1), also known as ATG1 (Autophagy related protein 1), GenBank Accession No. NM_003565, a.a. 2-end with N-terminal FLAG-tag, MW = 113 kDa, expressed in FreeStyle 293-F cells.

Complex of recombinant human CDK1 and CyclinA2, full length, with N-terminal GST-tags (GenBank Accession No. NM_001786, and NM_001237 respectively), co-expressed by Sf9 insect cells via a baculovirus expression system.

Recombinant full-length human CDK2 (GenBank Accession No. NM_001798), a.a. 2-298 (end) with N-terminal GST-tag, MW = 61 kDa, and CyclinA2 (GenBank Accession No. NM_001237), a.a. 174-432 (end) with N-terminal His-GST-tags, MW = 75 kDa. Both proteins were coexpressed in a baculovirus Sf9 insect cell expression system.

Recombinant human complex of CDK2 (cyclin dependent kinase 2), encompassing amino acids 2-298(end) and cyclinE1, encompassing amino acids 2-410(end). Both CDK2 and cyclinE1 constructs contain an N-terminal GST-tag, followed by a Thrombin Cleavage Site. The proteins were co-expressed in Sf9 cells and purified.

Complex of N-terminal GST-tagged recombinant full length human CDK3 (GenBank Accession No. NM_001258), and Cyclin E1 (GenBank Accession No. NM_001238). Co-expressed in a Baculovirus-infected Sf9 cell expression system. CDK3 MW=61 kDa, Cyclin E1 MW=74 kDa.