1. Antibodies
  2. Homo sapiens
  3. SOX13

ENCAB000BIR

Antibody against Homo sapiens SOX13

Homo sapiens
HepG2
characterized to standards with exemption
Status
released
Source (vendor)
Sigma
Product ID
WH0009580M1
Lot ID
10061-3E8
Characterized targets
SOX13 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Isotype
IgG1κ
Antigen description
SOX13 (NP_005677, a.a. 25-139) partial recomb protein w/ GST tag. MW of GST tag is 26 kDa.
External resources

Characterizations

SOX13 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
HepG2 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0009580M1). 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 HepG2: 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, SOX13, was identified as the 13th ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 SOX13-1.pdf
SOX13 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
not compliant
Caption
HepG2 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0009580M1). 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 2 from HepG2: 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, SOX13, was identified as the 37th ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
Reviewer comment
Not in the top 20 proteins by fold enrichment
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 SOX13-2.pdf
SOX13 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
not compliant
Caption
HepG2 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0009580M1). 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 3 from HepG2: 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, SOX13, was identified as the 62nd ranked enriched protein and the 2nd ranked transcription factor based on IP-Mass Spectrometry.
Reviewer comment
Not within top 20 proteins by fold enrichment
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 SOX13-3.pdf
SOX13 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
HepG2 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0009580M1). 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 4 from HepG2: 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, SOX13, was identified as the 7th ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 SOX13-4.pdf
SOX13 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not reviewed
Submitter comment
IP for Mass Spec data.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
SOX13 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Whole cell lysates of HepG2 were immunoprecipitated using the primary antibody (Sigma; WH0009580M1). 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. Four bands were detected at ~48, 57, 62, and 70-120 kDa.
Submitter comment
Rescued by mass spectrometry data. The size range of the bands is unexpected, but the overall analysis indicates that the antibody passes mass spec. SOX13 is known to heterodimerize with other proteins, including other TFs, but it is still the top TF in all mass spec analyses.
Reviewer comment
Expected size according to uniprot is ~69-70 kDa. The nearest band is not 50% of the overall signal. Missing IgG control.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 SOX13_IP-WB.png
SOX13 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
An immunoprecipitation followed by Western blot was performed with Sigma WH0004090M1 (lot 10061-3E8) antibody against SOX13 in HepG2 whole cell lysate along with a control IP with no lysate. The expected size of SOX13 is 98 kDa. Bands of ~48, 57, 62, and 70-120 kDa were detected in the test IP and not in the control IP. The experiment was repeated and the bands were analyzed by mass spec in the secondary validation.
Submitter comment
non specific bands and no igg control
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB