ENCAB000BKU

Homo sapiens

any cell type or tissue

characterized to standards

K562

Method: immunoblot

compliant

Caption: 0.25ug unmodified Recombinant Histone H3 (Millipore 14-494) and 30ug of K562 Whole Cell Lysate (Abcam, ab7911) were resolved by electrophoresis on a 4-12% acrylamide gel. 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% nonfat dry milk 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. Panel A: anti Histone H3 (Abcam ab1791), band of expected size (~17kDa) visualized in both unmodified recombinant histone H3 and WCL, at similar intensities.Panel B: Band of expected size (~17kDa) visualized in WCL, and cannot be detected in unmodified recombinant Histone H3. Antibody seems to be specific since no reactivity with nonhistone proteins or unmodified histones was detected
Submitted by: Noam Shoresh
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: H3K4me3 Diagenode pAb-003-50 lot A5051-001P_NL_WB.png
Documents: encode:Antibody_Characterization_ENCODE3_February2014.pdf

Method: dot blot assay

compliant

Caption: Performed dot blot analysis using a large set of modified peptides. The only peptide recognized by the H3K4me3 antibody was the correct one.
Submitted by: Meital Hatan
Lab: Bradley Bernstein, Broad
Grant: U54HG006991
Download: H3K4me3_Diagenode_pAb-003-050_A5051-001P_DotBlot.png
Documents: encode:Antibody_Characterization_ENCODE3_February2014.pdf