Apc7 Overexpression Lysate

• Reactivity: Hu

Apc7 Overexpression Lysate Summary

• Description:

  Apc7 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC219871
  
  Accession#: NM_016238
  
  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: ANAPC7

Applications/Dilutions

• 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: 63 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 Apc7 Overexpression Lysate

• anaphase promoting complex subunit 7
• anaphase-promoting complex subunit 7
• APC7Cyclosome subunit 7

Background

APC7 (anaphase-promoting complex subunit 7) is a member of the E3 enzyme family. This protein contains TPR repeats and has a molecular weight of approximately 63 kD. The APC7 protein is located in the nucleus during interphase and the centrosome during metaphase/anaphase. This protein probably recruits Cdh1 into the APC complex. The APC7 protein functions with other members of the APC complex as a multisubunit cell cycle ubiquitin ligase, and a regulator of sister chromatid separation by degrading securins. In addition, this protein functions in ubiquitin-dependent cyclin catabolism, metaphase/anaphase transition, and spindle elongation. The APC7 protein comprises one subunit of the anaphase promoting complex including APC1-8, and other probable complex proteins APC9-11, Cdc26, Mnd2, Swm1. The APC complex is inactivated by protein kinase A and is activated by CDC20 and Cdh1. The Poly6113 antibody has been shown to be useful for Western blotting of the human APC7 protein.

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.

Bioinformatics

• Gene Symbol: ANAPC7