ENCAB000AUE
Antibody against Homo sapiens NFE2
Homo sapiens
K562
characterized to standards with exemption
Homo sapiens
any cell type or tissue
partially characterized
- Status
- released
- Source (vendor)
- GeneTex
- Product ID
- GTX102698
- Lot ID
- 39946
- Characterized targets
- NFE2 (Homo sapiens)
- Host
- rabbit
- Clonality
- polyclonal
- Isotype
- IgG
- Antigen description
- Recombinant fgmt within amino acids 1 and 373 of NF-E2 (Uniprot ID#Q16621)
- External resources
Characterizations
NFE2 (Homo sapiens)
compliant
- Caption
- 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, NFE2, was identified as the 12th ranked enriched protein in band 1 and the 1st ranked transcription factor based on IP-Mass Spectrometry.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- NFE2-1.png
NFE2 (Homo sapiens)
not compliant
- Caption
- 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, NFE2, was identified as the 21st ranked enriched protein in band 2 and the 1st ranked transcription factor based on IP-Mass Spectrometry.
- Submitter comment
- This is the second band
- Reviewer comment
- This should be the better than the 20th and yet it is the 21st
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- NFE2-2.png
NFE2 (Homo sapiens)
K562
exempt from standards
- Caption
- Whole cell lysates of K562 were immunoprecipitated using the primary antibody (GeneTex; GTX102698). 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 ~35 and ~45kDa.
- Submitter comment
- this was rescued by a mass spec
- Reviewer comment
- So, this doesn't seem to have the right bands but the mass spec looks good
- Submitted by
- Flo Pauli-Behn
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- NFE2_IP-WB2.png
NFE2 (Homo sapiens)
not reviewed
- Caption
- K562 whole cell lysate was immunoprecipitated using the primary antibody (GeneTex; GTX102698). 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.
- Submitter comment
- The exact gel used for mass spec
- Reviewer comment
- This gel is just to show the bands used for the mass spec
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- NFE2_IP_MS_WB1.png