1. Antibodies
  2. Homo sapiens
  3. POU5F1

ENCAB000AKD

Antibody against Homo sapiens POU5F1

Homo sapiens
H1
characterized to standards
Homo sapiens
GM23338, bipolar neuron
characterized to standards with exemption
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-9081
Lot ID
F2609
Characterized targets
POU5F1 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Isotype
IgG
Antigen description
Epitope corresponding to amino acids 1-134 of Oct3/4 of human origin.

Characterizations

POU5F1 (Homo sapiens)
Method: motif enrichment
Attachment PDF Icon
compliant
Caption
The motif for target POU5F1 is represented by the attached position weight matrix (PWM) derived from file ENCFF983TJP. 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: 0.940575358452, Global enrichment Z-score: 8.29525057088, Positional bias Z-score: 11.1890506193, Peak rank bias Z-score: 7.24287450023, Enrichment rank: 1.0.
Submitted by
Aditi Narayanan
Lab
Richard Myers, HAIB
Download
 ENCAB000AKD.pdf
POU5F1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
IP followed by mass spectrometry: Briefly, human ES 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. Gel fragments corresponding to the bands indicated above in the western blot image were excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. There the samples were 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 9 of lower band, and hit 8 of upper band (shown in bold font).
Reviewer comment
Passes ENCODE2 and 3 standards, but document is not presented clearly.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB
POU5F1 (Homo sapiens)
GM23338bipolar neuron
Method: immunoblot
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
The ENCODE Binding Working Group finds for some valuable biosamples that recreating a primary on well characterized antibodies is not cost effective. Therefore, they allow exemption from standards for these samples.
Submitter comment
The lab is asking for an exemption for IPSCs and bipolar spindle neurons due to the lack of resources 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
POU5F1 (Homo sapiens)
H1
Method: immunoblot
Attachment from submitter
Attachment from submitters
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-9081 in whole cell lysate of human ES cells. POU5F1 bands are indicated.
Submitted by
Richard Myers
Lab
Richard Myers, HAIB