ENCAB000AHK

Homo sapiens

K562, HepG2, GM12878, SK-N-SH

characterized to standards with exemption

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 PVDF membrane using wet transfer. 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 slightly above the expected size (~131kDa) was detected, probably due to post translational modifications.
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: HDAC6_Bethyl_A301-341A_A301-341A-1_WB_AS.png
Documents: encode:Antibody_Characterization_ENCODE3_February2014.pdf
encode:Antibody_Characterization_ENCODE3_August2015.pdf

K562

Method: immunoblot

compliant

Method: ChIP-string comparison

exempt from standards

Submitter comment: These characterizations were done in ENCODE2, before the current characterization methods.
Reviewer comment: These are being exempted as prior to standards. The associated primaries look good.
Submitted by: Bradley Bernstein
Lab: Bradley Bernstein, Broad
Grant: U54HG004570
Download: human_Hdac6_A301-341A_validation_Bernstein.pdf
Documents: encode:ChIP_DNase_FAIRE_DNAme_v2_2011.pdf