ENCAB606JHO

Antibody against Homo sapiens ELF4

Homo sapiens
K562, MCF-7, HEK293T
characterized to standards with exemption
Homo sapiens
HepG2, GM12878
not characterized to standards
Status
released
Source (vendor)
Bethyl Labs
Product ID
A302-659A
Lot ID
1
Characterized targets
ELF4 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1250
External resources

Characterizations

ELF4 (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 A302-659A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 70.730
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ELF4 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2, using the antibody A302-659A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 70.730
Reviewer comment
Please explain the band ~ 90kd
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ELF4 (Homo sapiens)
HEK293T
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HEK293T, using the antibody A302-659A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 70.730
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
ELF4 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
exempt from standards
Caption
Immunoprecipitation of ELF4 from K562 cells using A302-659A. Lane 1: input nuclear lysate. Lane 2: material immunoprecipitated with A302-659A. Lane 3: material immunoprecipitated using control IgG. Band A and Band B were excised from gel and subject to analysis by mass spectrometry. The expected band size is 71 kDa.
Submitter comment
We should ask the ENCODE Antibody Review Panel to review.
Reviewer comment
Factor is also not detected in band B. On December 19, 2016, The ENCODE Antibody Review Panel exempted IP-MS characterizations in which the target TF is not the highest ranked TF if the factor(s) ranking higher are/belong to known pieces of the transcription and replication machinery (e.g. DNA and RNA polymerases, Topo 2, ORC, pre-RC, histones).
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
Download
ELF4 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
exempt from standards
Caption
IP followed by mass spectrometry: Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using A302-659A, 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
We should ask the ENCODE Antibody Review Panel to review
Reviewer comment
Factor is also not detected in band B. On December 19, 2016, The ENCODE Antibody Review Panel exempted IP-MS characterizations in which the target TF is not the highest ranked TF if the factor(s) ranking higher are/belong to known pieces of the transcription and replication machinery (e.g. DNA and RNA polymerases, Topo 2, ORC, pre-RC, histones).
Submitted by
Kathrina Onate
Lab
Michael Snyder, Stanford
ELF4 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody A302-659A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 70.730
Reviewer comment
Not the major band
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
ELF4 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878, using the antibody A302-659A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.Molecular Weight: 70.730
Reviewer comment
Multiple bands and marked band not >50% of the total signal in lane (it's also higher than expected size).
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford