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APE Overexpression Lysate

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Western Blot: APE1 Overexpression Lysate (Adult Normal) [NBP2-05771] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for APE1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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APE Overexpression Lysate Summary

Description

APE Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC213298

Accession#: NM_080649

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
APEX1

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
35.4 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 APE Overexpression Lysate

  • AP endonuclease 1
  • APE
  • APE1
  • APE-1
  • APEAPEX nuclease (multifunctional DNA repair enzyme)
  • APENAP endonuclease class I
  • APEX deoxyribonuclease (apurinic or apyrimidinic)
  • APEX nuclease (multifunctional DNA repair enzyme) 1
  • APEX nuclease
  • APEX1
  • Apurinic-apyrimidinic endonuclease 1
  • APXAP lyase
  • DNA-(apurinic or apyrimidinic site) lyase
  • EC 3.1
  • EC 4.2.99.18
  • HAP1apurinic/apyrimidinic (abasic) endonuclease
  • multifunctional DNA repair enzyme
  • protein REF-1
  • redox factor 1
  • Redox factor-1
  • REF1 apurinic/apyrimidinic exonuclease
  • REF-1

Background

DNA-(apurinic or apyrimidinic site) lyase, also known as APE1 or APEX1, is a multifunctional mammalian proteins responsible for the cellular response to oxidative stress. Specifically, APE-1 both functions in the DNA base excision repair pathway and regulates the redox of transcriptional factors.

For DNA repair, APE1 functions as a mammalian apurinic/apyrimidinic (AP) endodeoxyribonuclease responsible for the repair of AP sites in DNA lesions induced by oxidative and alkylating agents.

APEX1 also exerts a reversible nuclear redox activity to regulate the DNA binding affinity and the activity of transcriptional factors by regulating the redox status of their DNA-binding domains.

APE1 antibodies are useful for studies on both DNA repair and transcriptional regulation.

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 APEX1