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ChIPAb+ EED - ChIP Validated Antibody and Primer Set
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REFERENCES
Synergy of Eed and Tsix in the repression of Xist gene and X-chromosome inactivation.
Shibata, Shinwa, et al. (2008) EMBO J., 27: 1816-26 (2008)
The murine polycomb group protein Eed is required for global histone H3 lysine-27 methylation.
Montgomery, Nathan D, et al. (2005) Curr. Biol., 15: 942-7 (2005)
Different EZH2-containing complexes target methylation of histone H1 or nucleosomal histone H3.
Kuzmichev, Andrei, et al. (2004) Mol. Cell, 14: 183-93 (2004)
Transcriptional repression mediated by the human polycomb-group protein EED involves histone deacetylation.
van der Vlag, J and Otte, A P (1999) Nat. Genet., 23: 474-8 (1999)
Species Reactivity Key Applications Host Format Antibody Type
H, M, R, Op, Ch, Ca, B WB, IP Rabbit null Polyclonal Antibody
Description:
ChIPAb+ EED
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Trade Name:
Upstate (Millipore)
Product Overview:
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.
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Specificity:
Recognizes EED, Mr 62-70 kDa.
Molecular Weight:
~62-70 kDa
Epitope:
a.a. 429-441
Immunogen:
A synthetic peptide corresponding to a.a. 429-441 of human EED, conjugated to KLH.
Species Reactivity:
Human
Mouse
Rat
Opossum
Chicken
Canine
Bovine
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Species Reactivity Note:
Human, Mouse.
Predicted to react with rat, opossum, chicken, canine (dog), and bovine (cow) based on sequence homology.
Application Notes:
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).
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Control:
Includes negative control rabbit IgG antibody and primers specific for human HoxA2 upstream region.
Quality Assurance:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from Ntera2 cells (3 X 106 cell e |