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EED cell lysate

Human EED Baculovirus-Insect cells Overexpression Lysate

Gene Names
EED; HEED; WAIT1
Applications
Western Blot
Synonyms
EED; Human EED Baculovirus-Insect cells Overexpression Lysate; Human EED/Embryonic Ectoderm Development Insect Cell Lysate (WB positive control); embryonic ectoderm development; Human HEED Overexpression Lysate; Human WAIT1 Overexpression Lysate; EED cell lysate
Ordering
For Research Use Only!
Host
Baculovirus-Insect cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
678aa
Sequence
Met1-Arg441
Applicable Applications for EED cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the human EED (O75530-1) (Met 1-Arg 441) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for EED cell lysate
EED is essential for regulating the repressive histone modification, histone 3 lysine 27 tri-methylation (H3K27me3) at many developmental genes. EED depletion significantly impeded erythroid maturation. EED depletion impaired the generation of hematopoietic stem cells. The function of EED within polycomb repressive complex 2 (PRC2) is mediated by a complex network of protein-protein interactions. The targeted disruption of EED's reader function can lead to allosteric inhibition of PRC2 catalytic activity. Eed(Delta/Delta) HSPCs exhibited increased attachment to a major extracellular matrix component, fibronectin. Thus, EED deficiency increases proliferation on one side but promotes quiescence possibly by enhanced adhesion to the hematopoietic niche on the other, and these conflicting events would lead to abnormal differentiation and functional defect of Eed(Delta/Delta) HSPCs.

This Human EED overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of EED protein from the overexpression lysate was verified.
References
Ura H, et al. (2011) Eed/Sox2 regulatory loop controls ES cell self-renewal through histone methylation and acetylation. EMBO J. 30(11): 2190-204.Montgomery ND, et al. (2007) Molecular and functional mapping of EED motifs required for PRC2-dependent histone methylation. J Mol Biol. 374(5): 1145-57.Jin Q, et al. (2003) The protein phosphatase-1 (PP1) regulator, nuclear inhibitor of PP1 (NIPP1), interacts with the polycomb group protein, embryonic ectoderm development (EED), and functions as a transcriptional repressor. J Biol Chem. 278(33): 30677-85.Showell C, et al. (2002) Identification of putative interaction partners for the Xenopus Polycomb-group protein Xeed. Gene. 291(1-2): 95-104.Rinchik EM, et al. (1993) N-ethyl-N-nitrosourea-induced prenatally lethal mutations define at least two complementation groups within the embryonic ectoderm development (eed) locus in mouse chromosome 7. Mamm Genome. 4(7): 349-53.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
45,524 Da
NCBI Official Full Name
polycomb protein EED isoform a
NCBI Official Synonym Full Names
embryonic ectoderm development
NCBI Official Symbol
EED
NCBI Official Synonym Symbols
HEED; WAIT1
NCBI Protein Information
polycomb protein EED; WD protein associating with integrin cytoplasmic tails 1
UniProt Protein Name
Polycomb protein EED
Protein Family
UniProt Gene Name
EED
UniProt Synonym Gene Names
hEED; WAIT-1
UniProt Entry Name
EED_HUMAN

NCBI Description

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, Jul 2008]

Uniprot Description

EED: a member of the superfamily of Polycomb group (PcG) methyltransferase proteins with WD-40 repeats. PcG complexes mediate chromatin modifications that contribute to transcriptional silencing during development and carcinogenesis. A part of the Polycomb-repressive complex 2 (PRC2) along with EZH2, SUZ12, RBBP4 and RBBP7 and possibly AEBP2. Interacts with HDAC, HDAC2, histone H1 and YY1. PRC2 trimethylates histone H3 on K9 and K27, transcriptionally repressing target gene including HOXC8, HOXA9, MYT1 and CDKN2A. Appears to be overexpressed in breast and colon cancers. Inhibitors of PRC2-mediated gene repression are being investigated as a cancer therapy. EED and EZH2 are also implicated in the silencing of the inactive X chromosome. Three human isoforms produced by alternative splicing or initiation have been described, and additional isoforms may be produced by alternative initiation.

Protein type: Methyltransferase

Chromosomal Location of Human Ortholog: 11q14.2-q22.3

Cellular Component: nucleoplasm; sex chromatin; pronucleus; ESC/E(Z) complex; cytoplasm; nucleolus; nucleus

Molecular Function: histone methyltransferase activity; identical protein binding; protein binding; chromatin binding

Biological Process: histone methylation; genetic imprinting; transcription, DNA-dependent; negative regulation of gene expression, epigenetic; gene expression; negative regulation of transcription, DNA-dependent; regulation of gene expression, epigenetic

Research Articles on EED

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Product Notes

The EED eed (Catalog #AAA8114516) is a Cell Lysate produced from Baculovirus-Insect cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 678aa. AAA Biotech's EED can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the EED eed for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Arg44 1. It is sometimes possible for the material contained within the vial of "EED, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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