1. Antibodies
  2. Homo sapiens
  3. ETV6

ENCAB000ABD

Antibody against Homo sapiens ETV6

Homo sapiens
K562, GM12878
characterized to standards with exemption
Status
released
Source (vendor)
Sigma
Product ID
WH0002120M1
Lot ID
12089-3B10
Characterized targets
ETV6 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Isotype
IgG1κ
Antigen description
ETV6 (AAH43399, a.a. 1-453) full length recombinant protein with GST tag.
External resources

Characterizations

ETV6 (Homo sapiens)
K562GM12878
Method: immunoblot
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Whole cell lysates of K562 and GM12878 were immunoprecipitated using the primary antibody (Sigma; WH0002120M1). The IP fraction was separated on a 12% acrylamide gel with the Bio-Rad PROTEAN II xi system. After separation, the samples were transferred to a nitrocellulose membrane with an Invitrogen iBlot system. The membrane was probed with the primary antibody (same as that used for IP) and a secondary HRP-conjugated antibody. The resulting bands were visualized with SuperSignal West Femto Solution (Thermo Scientific). Protein Marker (PM) is labeled in kDa. Two bands were detected at ~50 and ~60kDa.
Submitter comment
No distinct band; however, this characterization is rescued by the mass spec data.
Reviewer comment
This antibody characterization is rescued by the Mass Spectrometry.
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB
ETV6 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not reviewed
Caption
GM12878 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0002120M1). The IP fraction was loaded on a 12% Bio-Rad TGX gel and separated with the Bio-Rad Tetra Cell system. A gel fragment (rectangle outline) corresponding to the band indicated on the Coomassie Blue stained gel image was excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility.
Submitter comment
Part of by the mass spec data- this characterization is simply a supplement to the mass spec, but missing IgG control and clear bands
Reviewer comment
Part of by the mass spec data- this characterization is simply a supplement to the mass spec, but lacks IgG control and clear bands
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 ETV6 IP.png
ETV6 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment from submitter
Attachment from submitters
compliant
Caption
Analysis of gel fragment from GM12878: The sample was analyzed on a LTQ XL Linear Ion Trap Mass Spectrometer by LC-ESI-MS/MS. Peptides were identified using SEQUEST tandem mass spectral analysis with probability based matching at p < 0.05. SEQUEST results were reported with ProteinProphet protXML Viewer (TPP v4.4 JETSTREAM) and filtered for a minimum probability of 0.9. All protein hits that met these criteria were reported, including common contaminants. Fold enrichment for each protein reported was determined using a custom script based on the FC-B score calculation from the reference Mellacheruvu et al., 2013. The CRAPome: a contaminant repository for affinity purification mass spectrometry data. Nat. Methods. 10(8):730-736. Doi:10.1038/nmeth.2557. The target protein, ETV6, was identified as the 4th ranked enriched protein and the 2nd ranked transcription factor based on IP-Mass Spectrometry.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB