1. Antibodies
  2. Homo sapiens
  3. EZH2

ENCAB000BKV

Antibody against Homo sapiens EZH2phosphoT487

Homo sapiens
K562, HeLa-S3, Raji
characterized to standards
Homo sapiens
DOHH2, KMS-11, HCT116, SK-N-SH, smooth muscle cell, MM.1S, OCI-LY3, GM23248, hepatocyte, PC-3, bipolar neuron, SU-DHL-6, Loucy, OCI-LY7, PC-9, MCF-7, neural progenitor cell, GM23338, Karpas-422
characterized to standards with exemption
Status
released
Source (vendor)
Abcam
Product ID
ab109398
Lot ID
YJ082421DS
Characterized targets
EZH2phosphoT487 (Homo sapiens)
Host
rabbit
Clonality
monoclonal
Aliases
bradley-bernstein:PchAb 919
External resources

Characterizations

EZH2phosphoT487 (Homo sapiens)
DOHH2KMS-11HCT116K562SK-N-SHsmooth muscle cellMM.1SOCI-LY3GM23248hepatocytePC-3bipolar neuronSU-DHL-6LoucyOCI-LY7PC-9MCF-7neural progenitor cellGM23338Karpas-422
Method: immunoblot
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
The antibody characterization review board chose to accept the several passing primaries in lieu of specific tests for each of these many cell lines.
Submitter comment
Many passing primaries exist.
Reviewer comment
The review board approves accepting the primaries that we have.
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad
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 exempted.png
EZH2phosphoT487 (Homo sapiens)
Method: ChIP-seq comparison
Attachment PDF Icon
compliant
Caption
This validation relies on the use of antibodies to a chromatin regulator and a functionally related histone modification in DOHH2 cells, and the demonstration that highly similar (>0.85) patterns of enrichment are obtained with each antibody. The first antibody is to phosphorylated EZH2 (PchAb 919, ENCAB000BKV), and the second antibody is to H3K27me3 previously characterized to Encode standards (PchAb 683, ENCAB000BLA).
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad
EZH2phosphoT487 (Homo sapiens)
K562
Method: immunoblot
Attachment from submitter
Attachment from submitters
compliant
Caption
1X10e6 cells equivalent of nuclear extract from K562 cells and a comparable amount of LP treated WCE HeLA cells, 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. a strong band of the expected size was detected (~85kDa) in the K562 WCL, this band appears much weaker in the treated cells, this allows us to reach the conclusion that the ab is specific for the phosphorylated variant.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
EZH2phosphoT487 (Homo sapiens)
HeLa-S3K562Raji
Method: immunoblot
Attachment PDF Icon
compliant
Caption
We performed a WB using 20ug of various nuclear extracts. Extracts were resolved by electrophoresis on a 4-12% Bis-Tris, 1X MES running buffer, iBlot transfer, nitrocellulose, P3 for 8 minutes, Blocked in TBST w 5% NFDM for 1 hour, Antibodies diluted in TBST w 5% NFDm, exposure in ChemiDoc.
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad