ENCAB169CDD

Antibody against Homo sapiens H3K27me3

Homo sapiens
any cell type or tissue
characterized to standards
Status
released
Source (vendor)
Diagenode
Product ID
C15410069-10
Lot ID
A1821_D
Host
rabbit
Clonality
polyclonal
Aliases
bradley-bernstein:PchAb 1383

Characterizations

H3K27me3 (Homo sapiens)
HeLa-S3
Method: immunoblot
compliant
Caption
We performed a WB using 20ug of HeLa Nuclear Lysate. Lysate was resolved by electrophoresis on a 4-12% bis-tris 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. Band of expected size (~17kDa) visualized in HeLa nuclear extract and cannot be detected in unmodified recombinant Histone H3. Antibody seems to be specific since no reactivity with non histone proteins or unmodified histones was detected.
Reviewer comment
Recombinant histone H3 is not included in the western blot but is included in the secondary dot blot, as allowed by the ENCODE antibody characterization standards.
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad
H3K27me3 (Homo sapiens)
Method: dot blot assay
compliant
Caption
We performed a DB analysis using the large set of modified peptides provided in the Active Motif MODified Histone Peptide Array, catalog #13005. The only peptide recognized by the H3K27me3 antibody was the correct one.
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad