1. Antibodies
  2. Homo sapiens
  3. ATF7

ENCAB000BMO

Antibody against Homo sapiens ATF7

Homo sapiens
GM12878, K562, HEK293T
characterized to standards
Homo sapiens
HepG2, MCF-7
characterized to standards with exemption
Homo sapiens
HeLa-S3, liver
not characterized to standards
Status
released
Source (vendor)
Sigma
Product ID
HPA003384
Lot ID
R04563
Characterized targets
ATF7 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
Antigen description
Cyclic AMP-dependent transcription factor ATF-7 recombinant protein epitope signature tag (PrEST)
Antigen sequence
LPGPPVQMPSVISLARPVSMVPNIPGIPGPPVNSSGSISPSGHPIPSEAKMRLKATLTHQVSSINGGCGMVVGTASTMVTARPEQSQILIQHPDAPSPAQPQVSPAQPTPSTGGRRRRTVDEDPDERRQRFLE
External resources

Characterizations

ATF7 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878, using the antibody HPA003384. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 53.0
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ATF7 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7, using the antibody HPA003384. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Submitter comment
Our mass spec analysis in HepG2 showed that ATF7 runs high and can be detected in the higher band, even though we marked the image with the band near the expected size.
Reviewer comment
Indicated band not >50% of all bands in IP lane. However, IP-MS analysis in HepG2 yielded a consistent banding pattern and ATF7 was found in the higher band, running higher than expected.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
ATF7 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2, using the antibody HPA003384. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Reviewer comment
Multiple bands and marked band not >50% of total signal in the lane.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
ATF7 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from HepG2 nuclear cell lysates using the antibody HPA003384, 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
In band A, none of the proteins with higher peptide count than ATF7 are categorized as sequence-specific TFs. Among the proteins with the same peptide count is DBIRD complex subunit ZNF326, which is involved in POLII transcript elongation, HP1BP3 is a histone-binding protein, and ATF is suggested to be a general inhibitor of the histone deacetylase HDAC1. In band B, none of the proteins detected have been shown to be sequence-specific TFs (including D6R9P3, ZC3H13, ZSCAN2, D6RF44).
Reviewer comment
The target TF is not found in band B but is found in A, the most abundant band. I think that we should just consider this to be a protein that runs larger than expected. I think this one should pass.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ATF7 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line HepG2 using the antibody HPA003384. 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: 53.0.
Submitter comment
We analyzed all immunoreactive bands by mass-spec and detect the TF of interest.
Reviewer comment
There are multiple immunoreactive bands, all analyzed by mass-spec. The target TF is detected in the major band but it seems to run at a higher than expected size.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ATF7 (Homo sapiens)
GM12878K562HepG2HeLa-S3MCF-7liver
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 : GM12878, K562, HepG2, HelaS3, MCF7 and Liver using the antibody HPA003384. Molecular mass: 52967 Da
Reviewer comment
Band is either too faint (Lanes 1 and 6) or is not 50% of signal (Llanes 3, 4, and 5). Lane 2 is compliant
Submitted by
Trupti Kawli
Lab
Michael Snyder, Stanford
ATF7 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
b) Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody HPA003384 .The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG. Expected size ~ 52 kDa.
Submitted by
Trupti Kawli
Lab
Michael Snyder, Stanford
ATF7 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878, using the antibody HPA003384. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Reviewer comment
Bands too faint to be sure.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
Download
 expt1035_4.jpg
ATF7 (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody HPA003384. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 53.0
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford