ENCAB757LUG

Antibody against Homo sapiens PHF8

Homo sapiens
K562, HepG2, GM12878, endothelial cell of umbilical vein, A549, H1
characterized to standards with exemption
Status
released
Source (vendor)
Millipore
Product ID
09-868
Lot ID
NRG1867761
Characterized targets
PHF8 (Homo sapiens)
Host
rabbit
Antigen description
PHD finger protein 8; jumonji C domain-containing histone lysine demethylase 1F
Aliases
bradley-bernstein:PchAb 1243
External resources

Characterizations

PHF8 (Homo sapiens)
Method: ChIP-seq comparison
exempt from standards
Caption
This validation relies on the use of antibodies to a chromatin regulator and a functionally related histone modification in A549 cells, and the demonstration that highly similar patterns of enrichment are obtained with each antibody. The first track shown used an antibody to PHF8 (PchAb 1243, ENCAB757LUG), and the second track shown used an antibody to H3K4me3 previously characterized to Encode standards (PchAb 100-V, ENCAB000BLF).
Submitter comment
The Antibody Review Board reviewed this document and the over all experiments to determine the quality of the data. 03-5-20018
Reviewer comment
For the PHF8 antibody https://www.encodeproject.org/antibodies/ENCAB757LUG/, I would be supportive of releasing the A549 and HepG2 datasets. Although the IDR was listed as borderline, this is a co-activator which is difficult to ChIP and will also have worse signal to noise than a DNA binding TF. The data looks reasonable and the primary western blots are good for both cell lines. The ChiP-comparison is reasonable; we wouldn’t expect all peaks to be in common between H3K4me3 and PHF8.
Submitted by
Nina Farrell
Lab
Bradley Bernstein, Broad
PHF8 (Homo sapiens)
A549H1
Method: immunoblot
Attachment from submitter
compliant
Caption
Nuclear lysates from A549 (10ug) and H1 (10ug) were loaded into a 4-12% Bis-Tris gel in 1X MOPS running buffer. After separation, the samples were transferred to a nitrocellulose membrane using iblot. Membrane was blocked for an hour in room temperature, with 5% BSA in TBS-T 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. Bands were detected at the expected size.
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad
PHF8 (Homo sapiens)
K562HepG2GM12878endothelial cell of umbilical vein
Method: immunoblot
Attachment from submitter
compliant
Submitted by
Noam Shoresh
Lab
Bradley Bernstein, Broad