ENCAB000ALC

Antibody against Homo sapiens SREBF1

Homo sapiens
HeLa-S3, HepG2
characterized to standards
Homo sapiens
K562, MCF-7, A549
characterized to standards with exemption
Homo sapiens
any cell type or tissue
partially characterized
Status
released
Source (vendor)
Santa Cruz Biotech
Product ID
sc-8984
Lot ID
10211
Characterized targets
SREBF1 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity

Characterizations

SREBF1 (Homo sapiens)
A549
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: A549, using the antibody sc-8984. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 121.675
Submitter comment
There are lower weight bands (below 75kDa) because Uniprot: [P36956] states that SREBF1 has post-translational modification. In the Golgi complex SREBPs are cleaved sequentially by site-1 and site-2 protease. Also, on wikipedia (https://en.wikipedia.org/wiki/Sterol_regulatory_element-binding_protein), we can see the figure which showed the cleavage; SREBP1 was cut into equally two parts. So 60kDa bands are half of the full size.
Reviewer comment
There are prominent immunoreactive bands of smaller than the expected size.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody sc-8984. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 121.675
Submitter comment
There are lower weight bands (below 75kDa) because Uniprot: [P36956] states that SREBF1 has post-translational modification. In the Golgi complex SREBPs are cleaved sequentially by site-1 and site-2 protease. Also, on wikipedia (https://en.wikipedia.org/wiki/Sterol_regulatory_element-binding_protein), we can see the figure which showed the cleavage; SREBP1 was cut into equally two parts. So 60kDa bands are half of the full size.
Reviewer comment
There are prominent immunoreactive bands of smaller than the expected size.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody sc-8984. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 121.675
Submitter comment
There are lower weight bands (below 75kDa) because Uniprot: [P36956] states that SREBF1 has post-translational modification. In the Golgi complex SREBPs are cleaved sequentially by site-1 and site-2 protease. Also, on wikipedia (https://en.wikipedia.org/wiki/Sterol_regulatory_element-binding_protein), we can see the figure which showed the cleavage; SREBP1 was cut into equally two parts. So 60kDa bands are half of the full size.
Reviewer comment
There are prominent immunoreactive bands of smaller than the expected size.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
Method: motif enrichment
compliant
Caption
The motif for target SREBF1 is represented by the attached position weight matrix (PWM) derived from [file name(s) here]. 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: 0.778440224 Global enrichment Z-score: 2.484651326 Positional bias Z-score: 1.886165924 Peak rank bias Z-score: 1.956377507 Enrichment rank: 1.0
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
Method: immunoprecipitation
Attachment from submitter
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody sc-8984. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 121.675
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
not reviewed
Caption
Immunoprecipitation of SREBP1 from HepG2 cells using sc-8984 antibody. Lane 1: input nuclear lysate, Lane 2: material immunoprecipitated with sc-8984, Lane 3: material immunoprecipitated using control IgG. Band A was excised from the gel and subjected to analysis by mass spectrometry. IP followed by masspectrometry: Briefly, protein was immunoprecipitated from HepG2 whole cell lysates using sc-8984, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGE Bis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was silver-stained, gel fragments corresponding to the bands indicated were excised and destained using the SilverSNAP Stain for Mass Spectrometry (Pierce). Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an LTQ- Orbitrap (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Protein false discovery rate < 1%, 2 peptides per protein minimum). We report 27proteins identified in band A, although 3 of these are also present in a control immunoprecipitation and are thus likely to present due to non-specific association with the IP matrix. Of the specifically immunoprecipitated proteins, SREBP1 is the most abundant (40 peptides). Based on these observations, this band is likely due to the presence of immunoprecipitated SREBP1 and sc-8984 meets the ENCODE standard for validation by this criterion.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
Method: immunoblot
not reviewed
Caption
Western blot of HelaS3 and HepG2 nuclear extracts using the anti SREBP1 (sc-8984) detects a single high molecular weight protein of about 125KD. This band was further analyzed by IP-Mass Spec.
Submitted by
Michael Snyder
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation of SREBP1 from HepG2 cells using sc-8984 antibody. Lane 1: input nuclear lysate, Lane 2: material immunoprecipitated with sc-8984, Lane 3: material immunoprecipitated using control IgG. Band A was excised from the gel and subjected to analysis by mass spectrometry.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
compliant
Caption
IP followed by mass spectrometry: Briefly, protein was immunoprecipitated from HepG2 whole cell lysates using sc-8984, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGE Bis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was silver-stained, gel fragments corresponding to the bands indicated were excised and destained using the SilverSNAP Stain for Mass Spectrometry (Pierce). Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an LTQ- Orbitrap (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Protein false discovery rate < 1%, 2 peptides per protein minimum). We report 27proteins identified in band A, although 3 of these are also present in a control immunoprecipitation and are thus likely to present due to non-specific association with the IP matrix. Of the specifically immunoprecipitated proteins, SREBP1 is the most abundant (40 peptides).
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
SREBF1 (Homo sapiens)
HepG2HeLa-S3
Method: immunoblot
Attachment from submitter
compliant
Caption
Western Blot analysis of nuclear extracts from HepG (lane 1) and HeLa-S3 (lane 2) using anti-SREBP1 antibody (sc-8984). Expected protein band is 125 kD.
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford