1. Antibodies
  2. Homo sapiens
  3. TCF4

ENCAB000ACQ

Antibody against Homo sapiens TCF4

Homo sapiens
K562
characterized to standards with exemption
Status
released
Source (vendor)
Sigma
Product ID
WH0006925M2
Lot ID
12262-1F6
Characterized targets
TCF4 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Isotype
IgG1κ
Antigen description
TCF4 (NP_005019, a.a. 1-366) full length recombinant protein with GST tag.
External resources

Characterizations

TCF4 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment from submitter
Attachment from submitters
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, TCF4, was not identified in gel fragment 1 based on IP-Mass Spectrometry.
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB
TCF4 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment from submitter
Attachment from submitters
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, TCF4, was the 11th ranked enriched protein and the 2nd ranked transcription factor in gel fragment 2 based on IP-Mass Spectrometry.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
TCF4 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment from submitter
Attachment from submitters
compliant
Caption
Analysis of gel fragment 3 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, TCF4, was the 7th ranked enriched protein and the 1st ranked transcription factor in gel fragment 3 based on IP-Mass Spectrometry.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
TCF4 (Homo sapiens)
K562
Method: immunoblot
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Whole cell lysates of K562 were immunoprecipitated using the primary antibody (Sigma; WH0006925M2). The IP fraction was separated on a 12% acrylamide gel with the Bio-Rad PROTEAN II xi system. After separation, the sample was 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. Three bands were detected at ~49, ~65, and ~85kDa.
Submitter comment
The non specific bands were individually mass spec'ed so the multiple bands are accounted for. There is no IgG control because this characterization was done before the requirement.
Reviewer comment
There bands to not reflect the size of the TF, but the mass spec shows that it is there.
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB
Download
 TCF4_IP-WB.png
TCF4 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not reviewed
Caption
K562 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0006925M2). 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
This image is to show which bands are used for the mass spec
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB