CDT1 Recombinant Rabbit Monoclonal Antibody [JA30-81]
Catalog# ET1704-67
CDT1 Recombinant Rabbit Monoclonal Antibody [JA30-81]
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WB
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IHC-P
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IF-Cell
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Human
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unconjugated
Overview
Product Name
CDT1 Recombinant Rabbit Monoclonal Antibody [JA30-81]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human CDT1 aa 497-546 / 546.
Species Reactivity
Human
Validated Applications
WB, IHC-P, IF-Cell
Target Molecular Weight
Predicted band size: 60 kDa
Positive Control
HCT 116 cell lysate, HT-29 cell lysate, Jurkat cell lysate, HEK-293 cell lysate, HT-29, human skin tissue.
Conjugation
unconjugated
Clone Number
JA30-81
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IHC-P
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1:25-1:50
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IF-Cell
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1:100
Target
Function
Human Cdt1 is a nuclear localizing replication initiation factor that is expressed only during the G1 and S phases of the cell cycle. In conjunction with Cdc18, Cdt1 is required to load the MCM protein Cdc21 onto chromatin at the end of mitosis which is necessary to initiate DNA replication. After S-phase onset, Cdt1 protein levels decrease and are barely detectable in cells in early S-phase or G2. However, Cdt1 mRNA is expressed in S-phase-arrested cells, and its levels do not change dramatically during the cell cycle, suggesting that proteolytic degradation rather than transcriptional controls ensure proper accumulation of Cdt1. Cdt1 can associate with the DNA replication inhibitor geminin, which is present in the S and G2 phases of the cell cycle. Inhibition of DNA replication by geminin in cell-free DNA replication extracts can be reversed by the addition of excess Cdt1. Geminin may be responsible for preventing inappropriate origin firing by targeting Cdt1.
Background References
暂无
Sequence Similarity
Belongs to the Cdt1 family.
Post-translational Modification
Two independent E3 ubiquitin ligase complexes, SCF(SKP2) and the DCX(DTL) complex, mediated CDT1 degradation in S phase. Ubiquitinated by the DCX(DTL) 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. Phosphorylation at Thr-29 by CDK2 targets CDT1 for ubiquitination by SCF(SKP2) E3 ubiquitin ligase and subsequent degradation. The interaction with GMNN protects it against ubiquitination. Deubiquitinated by USP37.; Phosphorylation by cyclin A-dependent kinases at Thr-29 targets CDT1 for ubiquitynation by SCF(SKP2) E3 ubiquitin ligase and subsequent degradation. Phosphorylated at Thr-29 by MAPK8/JNK1, which blocks replication licensing in response to stress. Binding to GMNN is not affected by phosphorylation.
Subcellular Location
Nucleus.
UNIPROT
Synonyms
CDT 1 antibody
cdt1 antibody
CDT1_HUMAN antibody
Chromatin licensing and DNA replication factor 1 antibody
DNA replication factor antibody
DNA replication factor Cdt1 antibody
Double parked antibody
Double parked Drosophila homolog of antibody
Double parked homolog antibody
DUP antibody
ExpandImages
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Western blot analysis of CDT1 on different lysates with Rabbit anti-CDT1 antibody (ET1704-67) at 1/2,000 dilution.
Lane 1: HCT 116 cell lysate
Lane 2: HT-29 cell lysate
Lane 3: Jurkat cell lysate
Lane 4: HEK-293 cell lysate
Lysates/proteins at 15 µg/Lane.
Predicted band size: 60 kDa
Observed band size: 70 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-67) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HT-29 cells labeling CDT1 with Rabbit anti-CDT1 antibody (ET1704-67) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CDT1 antibody (ET1704-67) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-CDT1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-67, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"