ENCAB741YHG

Homo sapiens

HepG2, endothelial cell of umbilical vein

characterized to standards

Homo sapiens

GM12878

characterized to standards with exemption

GM12878

Method: immunoblot

exempt from standards

Caption: This antibody was used before a characterization for GM12878 was performed. It has been reviewed and exempted by the antibody characterization board.
Submitter comment: The antibody has a conforming western only in HepG2 and HUVEC. A third dataset (https://www.encodeproject.org/experiments/ENCSR091BOQ/) used this antibody in GM12878. Release of that single dataset is blocked due to the lack of primary validation in that cell type. We would like an exception to allow the release of this one dataset. We do not anticipate further use of this antibody and have no remaining inventory.
Reviewer comment: PJF: I agree that this dataset should be released without requiring a western blot in GM12978 (using whatever term is appropriate such as “characterized to standard with exemption”).
Submitted by: Nina Farrell
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: exempted.png
Documents: encode:Histone_Modification_Antibody_Characterization_ENCODE3_Oct2016.pdf

Method: ChIP-seq comparison

compliant

Caption: This validation relies on the use of antibodies to different members of a known complex, and the demonstration that highly similar patterns of enrichment are obtained with each antibody. The second track shown used an antibody to EZH2 (PchAb 58-V), another member of the PRC2 complex. Overall correlation score: SUZ12 vs EZH2: 0.9439.
Submitted by: Kathrina Onate
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: SUZ12 CST 3737BF lot 4 SAV (1).pdf
Documents: encode:Histone_Modification_Antibody_Characterization_ENCODE3_Oct2016.pdf

HepG2endothelial cell of umbilical vein

Method: immunoblot

compliant

Caption: Nuclear lysates from HepG2 (10ug), HUVEC (10ug), were loaded into a 4-12% Bis-Tris gel in 1X MES running buffer. After separation, the samples were transferred to a nitrocellulose membrane using the iblot system. Membrane was blocked for an hour in room temperature, with 5% BSA in TBS-T and blotted with primary antibody in the appropriate concentration over night at 4c. Membrane was washed and blotted with secondary HRP-conjugated antibody. Detection was made with Optiblot ECL Detect Kit (ab133406) for 2 min.A band of the expected size was detected (~83kDa).
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: SUZ12_CellSignaling_3737BF_Lot4_WB.png
Documents: encode:Antibody_Characterization_ENCODE3_February2014.pdf