ENCAB000BKW

Antibody against Homo sapiens EZH2

Homo sapiens
K562, HEK293
characterized to standards
Homo sapiens
A673
characterized to standards with exemption
Status
released
Source (vendor)
Active Motif
Product ID
39639
Lot ID
23609001
Characterized targets
EZH2 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Aliases
bradley-bernstein:PchAb 150
External resources

Characterizations

EZH2 (Homo sapiens)
A673
Method: immunoblot
Attachment from submitter
exempt from standards
Caption
Both ENCODE submitter and antibody review board have agreed that this is a well-established antibody lot. Hence it does not require additional validation as standardized by the ENCODE Consortium.
Submitter comment
The lab requests that this is considered a well-documented antibody lot and therefore does not require additional validation.
Reviewer comment
Dr. Peggy J Farnham: I would support a “no additional validation needed” classification for this antibody.
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad
Download
EZH2 (Homo sapiens)
Method: ChIP-seq comparison
compliant
Caption
This validation relies on the use of antibodies to a chromatin regulator and a functionally related histone modification in A673 cells, and the demonstration that highly similar (>0.9) patterns of enrichment are obtained with each antibody. This antibody is to phosphorylated EZH2 (PchAb 150, ENCAB000BKW), and the second antibody is to H3K27me3 previously characterized to Encode standards (PchAb 848, ENCAB036YAO).
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad
EZH2 (Homo sapiens)
K562HEK293
Method: immunoblot
Attachment from submitter
compliant
Caption
5X10e5 cells equivalent of nuclear extract from HEK293 and K562 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. Band of the expected size was detected (~96kDa), the primary reactive band contains at least 50% of the signal observed on the blot. Submitting lab has confirmed with the vendor that lots 10310002,23609001 and 23809001 are identical.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad