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17-10034

Sigma-Aldrich

ChIPAb+ EED - ChIP Validated Antibody and Primer Set

from rabbit

Synonym(s):

EED, polycomb protein EED, WD protein associating with integrin cytoplasmic tails 1, WAIT-1

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.32

biological source

rabbit

Quality Level

clone

polyclonal

species reactivity

canine, mouse, human

species reactivity (predicted by homology)

opossum, dog, chicken, bovine, rat

manufacturer/tradename

ChIPAb+
Upstate®

technique(s)

ChIP: suitable
immunoprecipitation (IP): suitable
western blot: suitable

NCBI accession no.

UniProt accession no.

shipped in

dry ice

Gene Information

human ... EED(8726)

General description

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ EED, set includes the polycomb protein EED antibody, a negative control antibody (purified mouse IgG), and qPCR primers which amplify a 138 bp region upstream of human HoxA2 gene. The EED and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of EED associated chromatin.

Specificity

Recognizes EED, Mr 62-70 kDa.

Immunogen

A synthetic peptide corresponding to a.a. 429-441 of human EED, conjugated to KLH.
Epitope: a.a. 429-441

Application

Chromatin Immunoprecipitation:
Representative lot data.
Sonicated chromatin prepared from Ntera2 cells (3 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 µg of either a normal rabbit IgG or Anti-EED antibody and the Magna ChIP A Kit (Cat. # 17-610).
Successful immunoprecipitation of EED associated DNA fragments was verified by qPCR using GAPDH promoter (negative) and HoxA2 (positive) Primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each amplicon and ChIP sample as indicated.
Please refer to the EZ-Magna A ChIP (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Western Blot Analysis:
Representative lot data.
Lysates from Jurkat cell lysate were resolved by electrophoresis, transferred to PVDF and probed with anti-EED. Proteins were visualized using goat anti-rabbit secondary antibody conjugated to HRP and chemiluminescence detection (Please see figures).
This ChIPAb+ EED -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Quality

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from Ntera2 cells (3 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal rabbit IgG or Anti-EED antibody and the Magna ChIP® A Kit (Cat. # 17-610).
Successful immunoprecipitation of EED associated DNA fragments was verified by qPCR using Control Primers (Please see figures).

Please refer to the EZ-Magna ChIP A (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Target description

~62-70 kDa

Physical form

Anti-EED (Rabbit polyclonal IgG). One vial containing 50 μg of affinity purified antibody in 50 μL of 0.1 M Tris-Glycine (pH7.4) 150 mM NaCl, containing 0.05% azide. Store at -20°C.

Normal rabbit IgG. 1 vial containing 125 μg of purified rabbit IgG in 125 μL of storage buffer containing 0.1% sodium azide. Store at -20°C.

ChIP Primers, HoxA2 upstream. One vial containing 75 μL of 5 μM of each primer specific for the promoter region of human HoxA2. Store at -20°C.
FOR: AGG AAA GAT TTT GGT TGG GAA G
REV: AAA AAG AGG GAA AGG GAC AGA C
Format: Purified

Analysis Note

Control
Includes negative control rabbit IgG antibody and primers specific for human HoxA2 upstream region.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Storage Class Code

12 - Non Combustible Liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Andrei Kuzmichev et al.
Molecular cell, 14(2), 183-193 (2004-04-22)
Human Enhancer of Zeste homolog (Ezh2) is a histone lysine methyltransferase (HKMT) associated with transcriptional repression. Ezh2 is present in several distinct complexes, one of which, PRC2, we characterized previously. Here we report an additional Ezh2 complex, PRC3. We show
Nathan D Montgomery et al.
Current biology : CB, 15(10), 942-947 (2005-05-27)
PcG proteins mediate heritable transcriptional silencing by generating and recognizing covalent histone modifications. One conserved PcG complex, PRC2, is composed of several proteins including the histone methyltransferase (HMTase) Ezh2, the WD-repeat protein Eed, and the Zn-finger protein Suz12. Ezh2 methylates
Malay Mandal et al.
Nature immunology, 12(12), 1212-1220 (2011-11-01)
During B lymphopoiesis, recombination of the locus encoding the immunoglobulin κ-chain complex (Igk) requires expression of the precursor to the B cell antigen receptor (pre-BCR) and escape from signaling via the interleukin 7 receptor (IL-7R). By activating the transcription factor
Wensheng Zhang et al.
Cell stem cell, 24(1), 138-152 (2019-01-05)
BAF complexes are composed of different subunits with varying functional and developmental roles, although many subunits have not been examined in depth. Here we show that the Baf45 subunit Dpf2 maintains pluripotency and ESC differentiation potential. Dpf2 co-occupies enhancers with Oct4, Sox2
Shaghayegh Nouruzi et al.
Nature communications, 13(1), 2282-2282 (2022-04-29)
Treatment with androgen receptor pathway inhibitors (ARPIs) in prostate cancer leads to the emergence of resistant tumors characterized by lineage plasticity and differentiation toward neuroendocrine lineage. Here, we find that ARPIs induce a rapid epigenetic alteration mediated by large-scale chromatin

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