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Key Specifications Table
Species Reactivity | Key Applications |
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H, M | WB, ChIP |
Description | |
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Catalogue Number | 17-663 |
Brand Family | Upstate |
Trade Name |
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Description | ChIPAb+ EED - ChIP Validated Antibody and Primer Set |
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. |
Alternate Names |
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Product Information | |
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Format | Purified |
Control |
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Presentation | Anti-EED (Mouse monoclonal IgG). One vial containing 50 μg of protein G purified antibody in 0.1 M Tris-Glycine (pH7.4) 150 mM NaCl, containing 0.05% azide. May contain 30% glycerol (see certificate of analysis for details). Store at -20°C. Normal mouse IgG. Two vials containing 25 μg of purified mouse IgG in 25 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 |
Quality Level | MQ100 |
Applications | |
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Application | This ChIPAb+ EED -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
Key Applications |
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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 mouse IgG, or Anti-EED antibody and the Magna ChIP G Kit (Cat. # 17-611). 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 G ChIP™ (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. Western Blot Analysis: Representative lot data. Lysates from A431 cell lysate were resolved by electrophoresis, transferred to PVDF and probed with anti-EED. Proteins were visualized using goat anti-mouse secondary antibody conjugated to HRP and chemiluminescence detection (Please see figures). |
Biological Information | |
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Immunogen | Recombinant fusion protein of full length murine EED tagged with MBP. |
Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
Host | Mouse |
Specificity | Recognizes EED, MW: ~50 kDa. |
Isotype | IgG2aκ |
Species Reactivity |
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Species Reactivity Note | Human, Mouse. Not tested in other species. |
Antibody Type | Monoclonal Antibody |
Entrez Gene Number |
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Entrez Gene Summary | 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] |
Gene Symbol |
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UniProt Number |
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UniProt Summary | Function: Polycomb group (PcG) protein. Component of the PRC2/EED-EZH2 complex, which methylates Lys-9 and Lys-27 of histone H3, leading to transcriptional repression of the affected target gene. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1 and CDKN2A. Subunit Structure: Interacts with MLL By similarity. Component of the PRC2/EED-EZH2 complex, which includes EED, EZH2, SUZ12, RBBP4 and RBBP7 and possibly AEBP2. The minimum components required for methyltransferase activity of the PRC2/EED-EZH2 complex are EED, EZH2 and SUZ12. Component of the PRC2/EED-EZH1 complex, which includes EED, EZH1, SUZ12, RBBP4 and AEBP2. The PRC2 complex may also interact with DNMT1, DNMT3A, DNMT3B and PHF1 via the EZH2 subunit and with SIRT1 via the SUZ12 subunit. Interacts with HDAC, HDAC2, histone H1 and YY1. May interact with ITGA4, ITGAE and ITGB7. May interact with the MA protein of HIV-1. Subcellular Location: Nucleus. Note: Transiently colocalizes with XIST at inactive X chromosomes. Tissue Specificity: Expressed in brain, colon, heart, kidney, liver, lung, muscle, ovary, peripheral blood leukocytes, pancreas, placenta, prostate, spleen, small intestine, testis, thymus and uterus. Appears to be overexpressed in breast and colon cancer. Developmental Stage: Expression peaks at the G1/S phase boundary. Induction: Expression is induced by E2F1, E2F2 and E2F3. Sequence similarities: Belongs to the WD repeat ESC family. Contains 7 WD repeats. |
Molecular Weight | ~50 kDa |
Product Usage Statements | |
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Quality Assurance | 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 mouse IgG, or Anti-EED antibody and the Magna ChIP™ G Kit (Cat. # 17-611). Successful immunoprecipitation of EED-associated DNA fragments was verified by qPCR using Control Primers (Please see figures). Please refer to the EZ-Magna ChIP™ G (Cat. # 17-409) or EZ-ChIP™ (Cat. # 17-371) protocol for experimental details. |
Usage Statement |
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Storage and Shipping Information | |
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Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage. |
Packaging Information | |
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Material Size | 25 assays |
Material Package | 25 assays per set. Recommended use: ~2 μg antibody per chromatin immunoprecipitation (dependent upon biological context). |