Analysis Note

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

Application

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

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).

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

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

Epitope: a.a. 429-441

Legal Information

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

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

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

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.

Specificity

Recognizes EED, Mr 62-70 kDa.

Target description

~62-70 kDa