ENCAB000AEX

Homo sapiens

HeLa, K562, HepG2, SK-N-SH, A549, H1

characterized to standards

Homo sapiens

GM12878

not characterized to standards

Method: ChIP-seq comparison

compliant

Caption: This validation relies on the use of antibodies to chromatin regulators in K562 cells, and demonstrates highly similar patterns of enrichment are obtained with each antibody. The first track shown used an antibody to CBX2 (PchAb 165, ENCAB000AEV), and the second track shown used an antibody to CBX8 (PchAb 159, ENCAB000AEX). The third track shows data of a functionally related histone modification (H3K27me3) previously characterized to Encode standards (PchAb 67-V, ENCAB000ASB) – the use of this data is meant to act as a reference for the location in the genome rather than to quantitate to the track-track comparison between CBX2 and CBX8. Correlation of .947
Submitted by: Nina Farrell
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: CBX8 PchAb 159 SAV.pdf
Documents: encode:TF_Antibody_Characterization_ENCODE3_May2016.pdf

K562HepG2GM12878SK-N-SH

Method: immunoblot

compliant

Caption: Nuclear lysates from K562 (10ug), HepG2 (7ug), GM12878 (7ug), SK-N-SH (6ug), were loaded into a 4-12% Bis-Tris gel in 1X MOPS running buffer. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen 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 expected size was detected (~43kDa)
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: CBX8_Bethyl_A300-882A_A300-882A-1_WB.png
Documents: encode:Antibody_Characterization_ENCODE3_February2014.pdf
encode:Antibody_Characterization_ENCODE3_August2015.pdf

A549H1

Method: immunoblot

compliant

Caption: Nuclear lysates from A549 (10ug) and H1 (10ug) were loaded into a 4-12% Bis-Tris gel in 1X MOPS running buffer. After separation, the samples were transferred to a nitrocellulose membrane using iblot. 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. Bands were detected at 40kDa for H1 and at a range above the expected size (45kDa) for both cell types.
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: CBX8_Bethyl_A300-882A_A300-882A-1V2.png
Documents: encode:Antibody_Characterization_ENCODE3_August2015.pdf

Method: ChIP-string comparison

not reviewed

HeLa

Method: immunoblot

compliant