Not available outside of the UK & Ireland.

Analysis Note

ControlIncludes negative control mouse IgG antibody and primers specific for human HoxA2 upstream region.

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 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).

Research CategoryEpigenetics & Nuclear Function

Research Sub CategoryChromatin Biology

This ChIPAb+ EED -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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.

Immunogen

Recombinant fusion protein of full length murine EED tagged with MBP.

Legal Information

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

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

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Packaging

25 assays per set. Recommended use: ~2 µg antibody per chromatin immunoprecipitation (dependent upon biological context).

Physical form

Format: Purified

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

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.

Specificity

Recognizes EED, MW: ~50 kDa.

Not tested in other species.

Storage and Stability

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.

Target description

~50 kDa