1. Antibodies
  2. Homo sapiens
  3. NRF1

ENCAB000AJI

Antibody against Homo sapiens NRF1

Homo sapiens
GM12878, K562, HeLa-S3, MCF-7
characterized to standards
Homo sapiens
any cell type or tissue
partially characterized
Status
released
Source (vendor)
Abcam
Product ID
ab55744
Lot ID
GR27905
Characterized targets
NRF1 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Purification
Protein A/G
Antigen description
Immunogen corresponding to amino acids 201-286 of Human NRF1.

Characterizations

NRF1 (Homo sapiens)
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not submitted for review by lab
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody ab55744. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 54
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
NRF1 (Homo sapiens)
Method: motif enrichment
Attachment PDF Icon
compliant
Caption
The motif for target NRF1 is represented by the attached position weight matrix (PWM) derived from ENCFF227KGV. 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, as well as an enrichment rank. For more information on the underlying statistical methods, please see the attached document. Accept probability score: 0.919608216 Global enrichment Z-score: 7.203279708 Positional bias Z-score: 14.43478041 Peak rank bias Z-score: 13.37322462 Enrichment rank: 1.0
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
NRF1 (Homo sapiens)
Method: motif enrichment
Attachment PDF Icon
not reviewed
Caption
Calculations were done by Pouya Kheradpour using a collection of known motifs available at http://www.broadinstitute.org/%7Epouyak/motif-disc/human/. Table 1 shows the fold-enrichments, enrichment p-values and fraction of peaks which contain the motif. The motif which produced the largest value for each criterion is shown in Table 1. Motifs were identified using a matching stringency corresponding to 4-6 (6-mer). Peaks identified by IDR (1% cutoff) were used in the analysis and +/-50bp from peak centers were considered. Enrichments are for a given motif vs. a background consisting of +/- 50bp from the centers of all DnaseI hypersensitive peaks. Repeat mask/simple repeats from UCSC and all gencode v7 exons (including non-protein coding genes) were excluded from the analysis. Comparison to shuffle motifs were used to correct for compositional bias. Enrichment is the corrected # of motifs in ChIP peaks/corrected # of motifs in DNaseI peaks. The current ENCODE standard calls for >4-fold enrichment and >10% motif representation for this criteria to be used for validation. The 5 Nrf1 datasets presented here significantly exceed these thresholds and the antibody is considered validated. Motif consensus sequences and position weight matrices for highly enriched Nrf1 motif.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
NRF1 (Homo sapiens)
Method: immunoblot
Attachment PDF Icon
not reviewed
Caption
We observe one major band in all cell lines assayed that is approximately consistent with the size expected for NRF1 (55kD). Therefore, ab55744 meets this criterion for validation. Western blot using ab55744 against NRF1. Lanes contain nuclear lysates from GM12878 cells (Lane 1), K562 cells (Lane 2), and HeLa S3 cells (Lane 3).
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
NRF1 (Homo sapiens)
MCF-7
Method: immunoblot
Attachment from submitter
Attachment from submitters
compliant
Caption
Western blot analysis of nuclear lysates prepared from multiple cells lines loaded in the order: MCF7 using the antibody ab55744. Molecular Weight: 54
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
NRF1 (Homo sapiens)
GM12878K562HeLa-S3
Method: immunoblot
Attachment from submitter
Attachment from submitters
compliant
Caption
Western blot using ab55744 against NRF1. Lanes contain nuclear lysates from GM12878 cells (Lane 1), K562 cells (Lane 2), and HeLa-S3 cells (Lane 3). The expected band size for NRF1 for all cell lines is 55 kD.
Reviewer comment
Faint but consistent with expected size.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford