CDT1 Overexpression Lysate

Name: CDT1 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-09063

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-09063

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for CDT1 Overexpression Lysate

CDT1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC201657

Accession#: NM_030928

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

60.2 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.

Type

Overexpression

Scientific Data Images for CDT1 Overexpression Lysate

Western Blot: CDT1 Overexpression Lysate [NBL1-09063]

Western Blot: CDT1 Overexpression Lysate (Adult Normal) [NBL1-09063] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CDT1.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: CDT1

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.

Alternate Names

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

Gene Symbol

CDT1

Additional CDT1 Products

Product Documents for CDT1 Overexpression Lysate

Product Datasheets

Download Product Datasheets

COA

SDS

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Product Specific Notices for CDT1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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