ENCAB000AGE

Antibody against Homo sapiens ESRRA

Homo sapiens
MCF-7, K562, A549, HepG2
characterized to standards
Homo sapiens
any cell type or tissue
partially characterized
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-66882
Lot ID
A0809
Characterized targets
ESRRA (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
other
Isotype
IgG
Antigen description
Epitope corresponding to amino acids 81-160 mapping near the N-terminus of ERRalpha of human origin

Characterizations

ESRRA (Homo sapiens)
A549
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: A549, using the antibody sc-66882. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 53
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ESRRA (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody sc-66882. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 53
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ESRRA (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody sc-66882. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Reviewer comment
Same band in IgG lane.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
Download
ESRRA (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using the antibody sc-66882, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGEBis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was stained by ColloidialCoomassie G-250 stain, gel fragments corresponding to the bands indicated were excised. Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an Orbitrap Elite mass spectrometer (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Peptide false discovery rate < 1%, 2 unique peptides per protein minimum, mass error < 10 ppm).
Submitter comment
ESRRA acts as a site-specific transcription regulator and has been also shown to interact with estrogen and the transcripton factor TFIIB by direct protein-protein contact. http://www.genecards.org/cgi-bin/carddisp.pl?gene=ESRRA&keywords=esrra PUF60 is in association with FUBP1 regulates MYC transcription at the P2 promoter through the core-TFIIH basal transcription factor. Acts as a transcriptional repressor through the core-TFIIH basal transcription factor. http://www.genecards.org/cgi-bin/carddisp.pl?gene=PUF60&keywords=puf60. Both TFIIB and TFIIH are components of preinitiation complex (abbreviated PIC) which is a large complex of proteins that is necessary for the transcription of protein-coding genes in eukaryotes and archaea. The minimal PIC includes RNA polymerase II and six general transcription factors: TFIIA, TFIIB, TFIID, TFIIE, TFIIF, and TFIIH. https://en.wikipedia.org/wiki/Transcription_preinitiation_complex. PUF60 is 60KD, is in the ESRRA gel slice, but TFIIB (35KD) and TFIIH (ERCC3) (89KD) are not in the gel slice.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ESRRA (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody sc-66882. Lane 1: input nuclear lysate. Lane 2: material immunoprecipitated with antibody. Lane 3: material immunoprecipitated using control IgG. Marked bands were excised from gel and subjected to analysis by mass spectrometry. Target molecular weight: 53.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ESRRA (Homo sapiens)
Method: motif enrichment
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 ERRA dataset presented here exceeds these thresholds and the antibody is considered validated.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
ESRRA (Homo sapiens)
Method: immunoblot
not reviewed
Caption
A primary band with a mobility consistent with the size expected for ERRa (46kD; indicated by arrow) is identified by immunoblot with nuclear lysates from HepG2 cells. Therefore, sc-66882 meets this criterion for validation.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
ESRRA (Homo sapiens)
HepG2
Method: immunoblot
Attachment from submitter
compliant
Caption
Immunoblot using sc-66882 with nuclear extracts from HepG2 cell line. Expected band size is 46 kD, indicated by arrow.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford