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ZEB1 Overexpression Lysate

Images

 
Western Blot: ZEB1 Overexpression Lysate (Adult Normal) [NBL1-18010] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ZEB1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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ZEB1 Overexpression Lysate Summary

Description

ZEB1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC217704

Accession#: NM_030751

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
ZEB1

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
123.9 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 ZEB1 Overexpression Lysate

  • AREB6
  • AREB6MGC133261
  • BZP
  • delta-crystallin enhancer binding factor 1
  • DELTAEF1
  • FECD6
  • Negative regulator of IL2
  • NIL-2-A zinc finger protein
  • NIL2A
  • NIL-2-A
  • posterior polymorphous corneal dystrophy 3
  • PPCD3
  • TCF8
  • TCF-8
  • TCF8BZP
  • transcription factor 8 (represses interleukin 2 expression)
  • Transcription factor 8
  • ZEB
  • ZEB1
  • ZFHEP
  • ZFHX1A
  • zinc finger E-box binding homeobox 1
  • zinc finger E-box-binding homeobox 1
  • zinc finger homeodomain enhancer-binding protein

Background

ZEB1 encodes a zinc finger transcription factor that represses T-lymphocyte-specific IL2 gene (MIM 147680) expression by binding to a negative regulatory domain 100 nucleotides 5-prime of the IL2 transcription start site (Williams et al., 1991 [PubMed 1840704]).[supplied by OMIM]

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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Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for ZEB1 Lysate (NBL1-18010). (Showing 1 - 1 of 1 FAQ).

  1. In looking at the Zeb1 antibodies on your website, the molecular weights of Zeb1 detected by several antibodies are different. Why is this?
    • ZEB1 is a rather large protein and undergoes a lot of post translational modifications such as phosphorylation and glycosylation (please see: UniProt P37275). The reason band patterns are different in the images may be due to sample type and how highly modified the ZEB1 protein is that the antibody is detecting. The more post translational modifications the higher you will detect the protein.

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Bioinformatics

Gene Symbol ZEB1