The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at 2-5 µg/mg of lysate.
1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use at an assay dependent concentration.
1/1000 - 1/2500. Detects a band of approximately 63 kDa (predicted molecular weight: 63 kDa).
FunctionCooperates with CDC6 to promote the loading of the mini-chromosome maintenance complex onto chromatin to form the pre-replication complex necessary to initiate DNA replication. Binds DNA in a sequence-, strand-, and conformation-independent manner. Potential oncogene.
Sequence similaritiesBelongs to the Cdt1 family.
Developmental stagePresent during G1 and early S phase of the cell cycle. Degraded during the late S, G2, and M phases.
DomainThe PIP-box K+4 motif mediates both the interaction with PCNA and the recuitment of the DCX(DTL) complex: while the PIP-box interacts with PCNA, the presence of the K+4 submotif, recruits the DCX(DTL) complex, leading to its ubiquitination.
Post-translational modificationsUbiquitinated by the DCX(DTL) complex, also named CRL4(CDT2) complex, in response to DNA damage, leading to its degradation. Ubiquitination by the DCX(DTL) complex is necessary to ensure proper cell cycle regulation and is PCNA-dependent: interacts with PCNA via its PIP-box, while the presence of the containing the 'K+4' motif in the PIP box, recruit the DCX(DTL) complex, leading to its degradation. The interaction with GMNN protects it against ubiquitination. Phosphorylated by cyclin A-dependent kinases which results in the binding of CDT1 to the F-box protein SKP2 and subsequent degradation. Binding to GMNN is not affected by phosphorylation.
Hernández-Pérez S et al. USP37 deubiquitinates Cdt1 and contributes to regulate DNA replication. Mol OncolN/A:N/A (2016).
Read more (PubMed: 27296872) »
Grandy RA et al. Genome-Wide Studies Reveal that H3K4me3 Modification in Bivalent Genes Is Dynamically Regulated during the Pluripotent Cell Cycle and Stabilized upon Differentiation. Mol Cell Biol36:615-27 (2015).
Read more (PubMed: 26644406) »