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
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
- Grant
- U54HG006996
- Download
- 1108_05_ELF4_A302-659A.jpg
ELF4 (Homo sapiens)
HepG2
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
- Grant
- U54HG006996
- Download
- 1108_06_ELF4_A302-659A.jpg
ELF4 (Homo sapiens)
HEK293T
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
- Grant
- U54HG006996
- Download
- 1112_05_ELF4_A302-659A.jpg
ELF4 (Homo sapiens)
K562
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
- Grant
- U54HG006996
- Download
- ELF4.jpeg
ELF4 (Homo sapiens)
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
- Grant
- U54HG006996
- Download
- ELF4_A302-659A final.pdf
ELF4 (Homo sapiens)
K562
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
- Grant
- U54HG006996
- Download
- Expt1053_4-ELF4-A302-659A.jpg
ELF4 (Homo sapiens)
GM12878
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
- Grant
- U54HG006996
- Download
- expt_1107_3-ELF4-A302-659A.jpg