ENCAB854ATP
Antibody against Homo sapiens NR2F6
Homo sapiens
K562, HepG2
characterized to standards with exemption
- Status
- released
- Source (vendor)
- GeneTex
- Product ID
- GTX100255
- Lot ID
- 39568
- Characterized targets
- NR2F6 (Homo sapiens)
- Host
- rabbit
- Clonality
- polyclonal
- Purification
- affinity
- Isotype
- IgG
- Antigen description
- Synthetic peptide w/ sequence corresp to a region within amino acids 344 and 404 of NR2F6
- External resources
Characterizations
NR2F6 (Homo sapiens)
compliant
- Caption
- K562 whole cell lysate was immunoprecipitated using the primary antibody (Genetex; GTX100255). The IP fraction was loaded on a 12% Bio-Rad TGX gel and separated with the Bio-Rad Tetra Cell system. Gel fragments (rectangle outline) corresponding to the bands indicated on the Coomassie Blue stained gel image were excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. Analysis of gel fragment 1 from K562: 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, NR2F6, was identified as the 1st enriched protein and the 1st transcription factor based on IP-Mass Spectrometry.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- NR2F6-1.png
NR2F6 (Homo sapiens)
not compliant
- Caption
- K562 whole cell lysate was immunoprecipitated using the primary antibody (Genetex; GTX100255). The IP fraction was loaded on a 12% Bio-Rad TGX gel and separated with the Bio-Rad Tetra Cell system. Gel fragments (rectangle outline) corresponding to the bands indicated on the Coomassie Blue stained gel image were excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. Analysis of gel fragment 2 from K562: 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, NR2F6, was not identified by IP-Mass Spectrometry.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- NR2F6-2.png
NR2F6 (Homo sapiens)
K562
not submitted for review by lab
- Reviewer comment
- IP gel for mass spectrometry analysis
- Submitted by
- Richard Myers
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- NR2F6_IP-Commassie1.png
NR2F6 (Homo sapiens)
K562HepG2
exempt from standards
- Caption
- Whole cell lysates of K562 and HepG2 were immunoprecipitated using the primary antibody (Genetex; GTX100255). 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 ~44 and 80 kDa. Expected size ~42 kDa
- Submitter comment
- --
- Reviewer comment
- Band 1 is close to expected size, but not 50% of overall signal. Band 2 is not within 20% of expected size
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- NR2F6_IP-WB.png