CDT1 Antibody

Western Blot: CDT1 Antibody [NB100-2567] - Human B cells. KSHV infection protects CDT1 from hypoxia mediated degradation. Mock or KSHV infected cells were grown under normoxic or hypoxic condition followed by measuring CDT1 levels. Western blot image submitted by a verified customer review.

Immunohistochemistry-Paraffin: CDT1 Antibody [NB100-2567] - Human islet cell carcinoma. Antibody used at a dilution of 1:1000 (1ug/mL).

Western Blot: CDT1 Antibody [NB100-2567] - Whole cell lysate from HeLa (5, 15 and 50 ug for WB; 1 mg for IP, 20% of IP loaded) and 293T (T; 50 ug) cells. Affinity purified rabbit anti- CDT1 antibody (NB100-2567) used for WB at 1 ug/mL (A and B) and used for IP at 3 ug/mg lysate (B).

• Reactivity: Hu
• Applications: WB, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

CDT1 Antibody Summary

• Immunogen: The immunogen recognized by this antibody maps to a region between residue 500 and the C-terminus (residue 546) of human Chromatin Licensing and DNA Replication Factor 1 using the numbering given in entry NP_112190.1 (GeneID 81620).
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CDT1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 1:1000 - 1:5000
  • Immunohistochemistry-Paraffin 1:1000 - 1:5000
  • Immunoprecipitation 2-5 ug/mg lysate
  • Western Blot 1:1000 - 1:2500
• Application Notes: Epitope retrieval with citrate buffer pH 6.0 is recommended for FFPE tissue sections.
• Theoretical MW: 60.4 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  HeLa Whole Cell Lysate
  CDT1 Overexpression Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Tris-Citrate/Phosphate (pH 7 - 8)
• Preservative: 0.09% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for CDT1 Antibody

• chromatin licensing and DNA replication factor 1
• DNA replication factor Cdt1
• Double parked homolog
• Double parked, Drosophila, homolog of
• DUPRIS2

Background

Geminin is a protein that inhibits DNA replication by preventing the loading of MCM complex on chromatin. Human CDT1 is a 65kDa protein that contains 546 amino acids and coimmunoprecipitates with Geminin. Inhibition of DNA replication by Geminin in cell-free cDNA replication extracts can be reversed by the addition of excess CDT1. In the normal cell cycle, CDT1 is present only in G1 and S phases, whereas Geminin is present in S and G2 phases. Wohlschlegel et al. (2000) concluded that their results suggest that Geminin inhibits inappropriate origin firing by targeting CDT1. This antibody was raised by a genetic immunization technique. Genetic immunization can be used to generate antibodies by directly delivering antigen-coding DNA into the animal, rather than injecting a protein or peptide (Tang et al. PubMed: 1545867; Chambers and Johnston PubMed 12910245; Barry and Johnston PubMed: 9234514). The animals cells produce the protein, which stimulates the animals immune system to produce antibodies against that particular protein. A vector coding for a partial fusion protein was used for genetic immunisation of a mouse and the resulting serum was tested in Western blot against an E.coli lysate containing that partial fusion protein. Genetic immunization offers enormous advantages over the traditional protein-based immunization method. DNA is faster, cheaper and easier to produce and can be produced by standard techniques readily amenable to automation. Furthermore, the antibodies generated by genetic immunization are usually of superior quality with regard to specificity, affinity and recognizing the native protein.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CDT1 Antibody (NB100-2567)(5)

Publications using NB100-2567

• Singh RK, Lamplugh ZL, Lang F et al. KSHV-encoded LANA protects the cellular replication machinery from hypoxia induced degradation PLoS Pathog. 2019-09-01 [PMID: 31479497] (WB, Human)
• Hubbi ME, Kshitiz, Gilkes DMet al. A nontranscriptional role for HIF-1alpha as a direct inhibitor of DNA replication. Sci Signal 2013-02-12 [PMID: 23405012]
• Hubbi ME, Luo W, Baek JH et al. MCM proteins are negative regulators of hypoxia-inducible factor 1 Mol Cell 2011-06-10 [PMID: 21658608] (WB, Human)
• Kim Y, Starostina NG, Kipreos ET et al. The CRL4Cdt2 ubiquitin ligase targets the degradation of p21Cip1 to control replication licensing. Genes Dev 2008-09-01 [PMID: 18794348]
• Pos Z, Wiener Z, Pocza P et al. Histamine suppresses fibulin-5 and insulin-like growth factor-II receptor expression in melanoma. Cancer Res 2008-03-01 [PMID: 18339882]

Reviews for CDT1 Antibody (NB100-2567) (3)

Average Rating: 3.3

(Based on 3 reviews)

We have 3 reviews tested in 1 species: Human.

reviewed by: Verified Customer WB Human 10/16/2019

Summary

• Application: Western Blot
• Sample Tested: B cells
• Species: Human
• Lot: A1

reviewed by: Nalin Ratnayeke ICC Human 01/17/2018

Summary

• Application: Immunocytochemistry
• Sample Tested: hela cell
• Species: Human
• Lot: A1

Comments

• Comments: Fixed in 4% PFA for 10 min, permeablized in .2% Triton X-100 in PBS for 15 min, blocked for 1 hr in 1% BSA + 10% FBS in PBS with .1% Triton X-100. Incubated in antibody at 1:100 dilution in blocking buffer, and detected with anti-rabbit secondary antibody.

reviewed by: Verified Customer WB Human 02/18/2011

Summary

• Application: Western Blot
• Sample Tested: HCT116 Whole Cell Lysate, Sample Amount: 50 ug
• Species: Human

Bioinformatics

• Gene Symbol: CDT1
• Entrez:
  • 81620(Human)
• Uniprot:
  • Q9H211(Human)