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Cdc27 Overexpression Lysate

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Western Blot: Cdc27 Overexpression Lysate (Adult Normal) [NBP2-09390] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Cdc27.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Cdc27 Overexpression Lysate Summary

Description

Cdc27 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC226198

Accession#: NM_001114091

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.
Gene
CDC27

Applications/Dilutions

Dilutions
  • Western Blot
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.
Theoretical MW
92.4 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.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

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.

Alternate Names for Cdc27 Overexpression Lysate

  • ANAPC3CDC27 homolog
  • anaphase promoting complex subunit 3
  • Anaphase-promoting complex subunit 3
  • anaphase-promoting complex, protein 3
  • APC 3
  • APC3cell division cycle 27
  • CDC27Hs
  • cell division cycle 27 homolog (S. cerevisiae)
  • cell division cycle protein 27 homolog
  • D0S1430E
  • D17S978E
  • HNUC
  • H-NUC
  • nuc2 homolog
  • NUC2

Background

Cdc27 shares strong similarity with Saccharomyces cerevisiae protein Cdc27, and the gene product of Schizosaccharomyces pombe nuc 2. It is a component of the Anaphase Promoting Complex (APC), which is composed of eight protein subunits and is highly conserved in eucaryotic cells. The APC catalyzes the formation of the cyclin B ubiquitin conjugate that is responsible for the ubiquitin mediated proteolysis of B type cyclins. This protein and 3 other members of the APC complex contain the TPR (tetratricopeptide repeat), a protein domain important for protein protein interaction. This protein was shown to interact with mitotic checkpoint proteins including Mad2, p55CDC and BUBR1, and thus may be involved in controlling the timing of mitosis.

Limitations

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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Bioinformatics

Gene Symbol CDC27