ENCAB000AFH

Homo sapiens

K562, HepG2, GM12878, endothelial cell of umbilical vein

partially characterized

K562

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. UniProt has 220 KD max, gel says to expect 260 KD.
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: chd4_bethyl_A301-081-1_IP.png
Documents: encode:Antibody_Characterization_ENCODE3_February2014.pdf
encode:Antibody_Characterization_ENCODE3_August2015.pdf

K562HepG2GM12878endothelial cell of umbilical vein

Method: immunoblot

compliant

Caption: Nuclear lysates from K562 (10ug), HepG2 (10ug), GM12878 (7ug), HUVEC (10ug), 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 single band at the expected size (~270kDa) was detected.
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: CHD4_Mi2.beta_Bethyl_A301-081A-1_WB.png
Documents: encode:Antibody_Characterization_ENCODE3_February2014.pdf
encode:Antibody_Characterization_ENCODE3_August2015.pdf