1. Antibodies
  2. Homo sapiens
  3. HNF4G

ENCAB000AHQ

Antibody against Homo sapiens HNF4G

Homo sapiens
liver
characterized to standards with exemption
Homo sapiens
HepG2
not characterized to standards
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-6558
Lot ID
I299
Characterized targets
HNF4G (Homo sapiens)
Host
goat
Clonality
polyclonal
Isotype
IgG
Antigen description
Epitope mapping at the C-terminus of HNF4G of human origin.

Characterizations

HNF4G (Homo sapiens)
Method: motif enrichment
Attachment PDF Icon
compliant
Caption
The motif for target HNF4G is represented by the attached position weight matrix (PWM) derived from file ENCFF313GAK. Motif enrichment analysis was done by Dr. Zhizhuo Zhang (Broad Institute, Kellis Lab) using known motifs (http://compbio.mit.edu/encode-motifs/) and previously published ChIP-seq data (http://www.broadinstitute.org/~zzhang/motifpipeline/data/TrainSetInfo.txt). The accept probability score of the given transcription factor was calculated using a Bayesian approach. This analysis also includes three motif enrichment scores, computed by overlapping the motif instances with the given ChIP-seq peak locations. For more information on the underlying statistical methods, please see the attached document. From ENCFF313GAK: Accept probability score: 0.803041780615, Global enrichment Z-score: 5.78518888723, Positional bias Z-score: 7.65798453867 , Peak rank bias Z-score: 5.61254638223. Enrichment rank: 1.0
Submitted by
Aditi Narayanan
Lab
Richard Myers, HAIB
Download
 ENCAB000AHQ.pdf
HNF4G (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
not compliant
Caption
Briefly, HepG2 whole cell lysates were immunoprecipitated using primary antibody, and the IP fraction was loaded on a 12% acrylamide gel and separated with a Bio-Rad PROTEAN II xi system. Gel was stained with Coomassie Blue in order to visualize marker bands. A gel fragment corresponding to the band indicated above in the western blot image was excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. There the sample was run on an LTQ XL Linear Ion Trap Mass Spectrometer by LC-ESI-MS/MS. Peptides were identified using SEQUEST tandem mass spectra analysis, with probability based matching at p < 0.05. As per ENCODE data standards, all SEQUEST results are attached, including common contaminants. Target protein is listed as hit 21a. Results in document "ENCODE_HAIB_HNF4G_(SC-6558)_03172011_MassSpec.pdf"
Reviewer comment
can't find data for this
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
HNF4G (Homo sapiens)
HepG2
Method: immunoblot
Attachment from submitter
Attachment from submitters
not compliant
Caption
Western blot protocol: Whole cell lysate was immunoprecipitated using primary antibody, and the IP fraction was loaded on a 12% acrylamide gel and separated with a Bio-Rad PROTEAN II xi system. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen iBlot system. Blotting with primary (same as that used for IP) and secondary HRP-conjugated antibodies was performed on an Invitrogen BenchPro 4100 system. Visualization was achieved using SuperSignal West Femto solution (Thermo Scientific). Results: Band of expected size visualized, representing strongest signal in the lane. Figure legend: IP-western with sc-6558 in HepG2 whole cell lysate (WCL); PM=protein marker. HNF4G band is indicated.
Reviewer comment
Not 50% of signal in lane
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
HNF4G (Homo sapiens)
liver
Method: immunoblot
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
The ENCODE Binding Working Group finds for some valuable tissues that recreating a primary on well characterized antibodies is not cost effective. Therefore, they allow exemption from standards for these tissues.
Submitter comment
The lab is asking for an exemption for liver cells due to the lack of resource to make a primary characterization for them
Reviewer comment
Exempted by the Feb 29, 2016 antibody review panel
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
Download
 No_tissue.png