ENCAB000AJV

Homo sapiens

HeLa

partially characterized

Homo sapiens

A549, H1, K562

not characterized to standards

Method: ChIP-string comparison

not reviewed

HeLa

Method: immunoblot

compliant

A549H1

Method: immunoblot

not compliant

Caption: Nuclear lysates from A549 (10ug) and H1 (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 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 in a range below the expected size (110kDa).
Reviewer comment: Band intensity is not 50% of total signal for both lanes
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: KAT2B_PCAF_Bethyl_A301-666A_A301-666A-1_V2.png
Documents: encode:Antibody_Characterization_ENCODE3_August2015.pdf

K562K562

Method: immunoprecipitation

not compliant

Caption: K562 whole cell lysate was immunoprecipitated using primary antibody, and the IP fraction along side a whole cell lysate were loaded on a CriterionXT gel and separated. After separation, the samples were transferred using a wet transfer. Blotting with primary (same as that used for IP) and secondary HRP-conjugated antibodies was performed. Band of expected size visualized representing strongest signal in the lane.
Reviewer comment: No IgG control
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: pcaf_bethyl_A300-666A-1_IP.png
Documents: encode:Antibody_Characterization_ENCODE3_August2015.pdf