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E6AP/UBE3A Overexpression Lysate

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Western Blot: UBE3A Overexpression Lysate (Adult Normal) [NBL1-17551] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for UBE3A.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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E6AP/UBE3A Overexpression Lysate Summary

Description

E6AP/UBE3A Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC216727

Accession#: NM_000462

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
UBE3A

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
100.5 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 E6AP/UBE3A Overexpression Lysate

  • ANCR
  • ANCREPVE6AP
  • AS
  • CTCL tumor antigen se37-2
  • E6AP ubiquitin-protein ligase
  • E6AP
  • E6-AP
  • EC 6.3.2.-
  • EPVE6AP
  • FLJ26981
  • HPVE6A
  • human papilloma virus E6-associated protein
  • Human papillomavirus E6-associated protein
  • Oncogenic protein-associated protein E6-AP
  • Renal carcinoma antigen NY-REN-54
  • UBE3A
  • ubiquitin protein ligase E3A
  • ubiquitin-protein ligase E3A

Background

This gene encodes an E3 ubiquitin-protein ligase, part of the ubiquitin protein degradation system. This imprinted gene is maternally expressed in brain and biallelically expressed in other tissues. Maternally inherited deletion of this gene causes Angelman Syndrome, characterized by severe motor and intellectual retardation, ataxia, hypotonia, epilepsy, absence of speech, and characteristic facies. The protein also interacts with the E6 protein of human papillomavirus types 16 and 18, resulting in ubiquitination and proteolysis of tumor protein p53. Alternative splicing of this gene results in three transcript variants encoding three isoforms with different N-termini. Additional transcript variants have been described, but their full length nature has not been determined. [provided by RefSeq]. Transcript Variant: This variant (2) encodes the longest isoform of this protein. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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 UBE3A