1. Antibodies
  2. Homo sapiens
  3. GATA4

ENCAB000BPU

Antibody against Homo sapiens GATA4

Homo sapiens
HepG2
characterized to standards with exemption
Status
released
Source (vendor)
Active Motif
Product ID
39894
Lot ID
26310001
Characterized targets
GATA4 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
Antigen description
antibody was raised against a peptide derived from the C-terminus of human GATA-4.
External resources

Characterizations

GATA4 (Homo sapiens)
Method: motif enrichment
Attachment PDF Icon
compliant
Caption
The motif for target GATA4 is represented by the attached position weight matrix (PWM) derived from file ENCFF013SXI. 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. Accept probability score: 0.818708558334, Global enrichment Z-score: 2.74307442199, Positional bias Z-score: 4.79675328826, Peak rank bias Z-score: 4.00530896457, Enrichment rank: 2.0.
Submitted by
Aditi Narayanan
Lab
Richard Myers, HAIB
Download
 ENCAB000BPU.pdf
GATA4 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment from submitter
Attachment from submitters
compliant
Caption
HepG2 whole cell lysate was immunoprecipitated using the primary antibody (Active Motif; 39894). The IP fraction was loaded on a 12% Bio-Rad TGX gel and separated with the Bio-Rad Tetra Cell system. A gel fragment (rectangle outline) corresponding to the band indicated on the Coomassie Blue stained gel image was excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. Analysis of gel fragment 1 from HepG2: The sample was analyzed on a LTQ XL Linear Ion Trap Mass Spectrometer by LC-ESI-MS/MS. Peptides were identified using SEQUEST tandem mass spectral analysis with probability based matching at p < 0.05. SEQUEST results were reported with ProteinProphet protXML Viewer (TPP v4.4 JETSTREAM) and filtered for a minimum probability of 0.9. All protein hits that met these criteria were reported, including common contaminants. Fold enrichment for each protein reported was determined using a custom script based on the FC-B score calculation from the reference Mellacheruvu et al., 2013. The CRAPome: a contaminant repository for affinity purification mass spectrometry data. Nat. Methods. 10(8):730-736. Doi:10.1038/nmeth.2557. The target protein, GATA4, was identified as the 10th ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 GATA4-1.png
GATA4 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Whole cell lysates of HepG2 were immunoprecipitated using the primary antibody (Active Motif; 39894). The IP fraction was separated on a 12% acrylamide gel with the Bio-Rad PROTEAN II xi system. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen iBlot system. The membrane was probed with the primary antibody (same as that used for IP) and a secondary HRP-conjugated antibody. The resulting bands were visualized with SuperSignal West Femto Solution (Thermo Scientific). Protein Marker (PM) is labeled in kDa. One band was detected at ~52 kDa. Expected size ~44-45 kDa.
Submitter comment
--
Reviewer comment
Missing IgG Control and the existing control also has the band of interest. Rescued by mass spectrometry data.
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB
GATA4 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not reviewed
Reviewer comment
IP gel for mass spectrometry analysis
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB