ENCAB036YAO

Antibody against Homo sapiens H3K27me3

Homo sapiens
any cell type or tissue
characterized to standards
Status
released
Source (vendor)
Millipore
Product ID
07-449
Lot ID
2120130
Characterized targets
H3K27me3 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Aliases
bradley-bernstein:PchAb 848
External resources

Characterizations

H3K27me3 (Homo sapiens)
Method: dot blot assay
Attachment from submitter
compliant
Caption
Performed dot blot analysis using a large set of modified peptides. The only peptide recognized by the H3K27me3 antibody was the correct one.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
H3K27me3 (Homo sapiens)
K562GM12878endothelial cell of umbilical vein
Method: immunoblot
Attachment from submitter
compliant
Caption
0.7ug unmodified Recombinant Histone H3 (Millipore 14-494) and 10ug of K562 Nuclear Lysate, were 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. 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 Whole cell lysates of K562, GM12878 and Huvec 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. according to the vendor lot numbers JBC1873477,JBC1854858 and 2039786 are from the same parent batch.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
H3K27me3 (Homo sapiens)
K562HEK293
Method: immunoblot
Attachment from submitter
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. according to the vendor lot numbers JBC1873477,JBC1854858 and 2039786 are from the same parent batch.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad