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
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
- Grant
- U54HG006998
- Download
- ETV6 immunoblot.png
ETV6 (Homo sapiens)
GM12878
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
- Grant
- U54HG006998
- Download
- ETV6 IP.png
ETV6 (Homo sapiens)
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
- Grant
- U54HG006998
- Download
- ETV6 Mass spec.png