ENCAB000AUF
Antibody against Homo sapiens TARDBP
Homo sapiens
K562, GM12878
characterized to standards
Homo sapiens
HepG2
characterized to standards with exemption
Homo sapiens
any cell type or tissue
partially characterized
- Status
- released
- Source (vendor)
- GeneTex
- Product ID
- GTX114210
- Lot ID
- 40135
- Characterized targets
- TARDBP (Homo sapiens)
- Host
- rabbit
- Clonality
- polyclonal
- Antigen description
- Recombinant fgmt within amino acids 1 and 289 of TARDBP (Uniprot ID#Q13148)
- External resources
Characterizations
TARDBP (Homo sapiens)
not 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, TARDBP, was not identified as an enriched protein in band 1 based on IP-Mass Spectrometry.
- Reviewer comment
- TARDBP does not show up in the list at all
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- TARDBP-1.png
TARDBP (Homo sapiens)
exempt from standards
- 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, TARDBP, was identified as the 27th enriched protein and the 2nd ranked transcription factor in band 2 based on IP-Mass Spectrometry.
- Submitter comment
- Although this is the 27th ranked peptide it is the 2nd ranked transcription factor
- Reviewer comment
- As per the decisions of the Antibody Review Panel on Feb 29, 2016. This characterization is exempted from standards.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- TARDBP-2.png
TARDBP (Homo sapiens)
GM12878
not submitted for review by lab
- Reviewer comment
- IP gel for mass spectrometry analysis
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- TARDBP_GM12878_IP-Coomassie_Gel.png
TARDBP (Homo sapiens)
compliant
- Caption
- GM12878 whole cell lysate was immunoprecipitated using the primary antibody (GeneTex; GTX114210). 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. Analysis of gel fragment 1 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, TARDBP, was identified as the 1st enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- TARDBP_GM12878_IP-MS.png
TARDBP (Homo sapiens)
K562GM12878
compliant
- Caption
- Whole cell lysates of K562 and GM12878 were immunoprecipitated using the primary antibody (GeneTex; GTX114210). 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. One band was detected at ~42 kDa.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- TARDBP_GM12878_IP-WB.png
TARDBP (Homo sapiens)
not reviewed
- Caption
- K562 whole cell lysate was immunoprecipitated using the primary antibody (GeneTex; GTX114210). 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
- This image is to show which bands are used for the mass spec
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- TARDBP_IP_MS_WB_1.png
TARDBP (Homo sapiens)
K562HepG2
exempt from standards
- Caption
- Whole cell lysates of K562 and HepG2 were immunoprecipitated using the primary antibody (GeneTex; GTX114210). 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 ~45 kDa.
- Submitter comment
- These characterizations were done during ENCODE2 when there was no requirement for IgG control.
- Reviewer comment
- As per Antibody review panal decsion of Feb 29, 2016, this will be exempted from standards
- Submitted by
- Flo Pauli-Behn
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- TARDBP_IP_WB.png