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EED Overexpression Lysate

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Western Blot: EED Overexpression Lysate (Adult Normal) [NBL1-10119] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for EED.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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EED Overexpression Lysate Summary

Description

EED Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC219261

Accession#: NM_003797

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
EED

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
50 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 EED Overexpression Lysate

  • EED
  • embryonic ectoderm development
  • HEED
  • polycomb protein EED
  • WAIT1
  • WAIT1
  • WAIT-1
  • WD protein associating with integrin cytoplasmic tails 1

Background

This gene encodes a member of the Polycomb-group (PcG) family. PcG family members form multimeric protein complexes, which are involved in maintaining the transcriptional repressive state of genes over successive cell generations. This protein interacts with enhancer of zeste 2, the cytoplasmic tail of integrin beta7, immunodeficiency virus type 1 (HIV-1) MA protein, and histone deacetylase proteins. This protein mediates repression of gene activity through histone deacetylation, and may act as a specific regulator of integrin function. Two transcript variants encoding distinct isoforms have been identified for this gene. [provided by RefSeq]. Transcript Variant: This variant (1) encodes the longer isoform (a). 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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Publications for EED Lysate (NBL1-10119) (0)

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Product General Protocols

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Video Protocols

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Research Areas for EED Lysate (NBL1-10119)

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Blogs on EED.

Microglia: pruning shears for homeostatic maintenance in the brain
By Jennifer Sokolowski, MD, PhD.Microglia play a critical role in pruning neurons and synapses during homeostatic maintenance in the adult brain.1 A recent study by Ayata et al. (2018) identified regional differe...  Read full blog post.

EZH1 has more to offer than gene repression
EZH1 is part of the Polycomb-group family of proteins, which are responsible for remodeling chromatin in genes and modulating epigenetic silencing during development.  Specifically, EZHI is a component of PRC2, or polycomb repressive complex-2.  PR...  Read full blog post.

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Bioinformatics

Gene Symbol EED