1. Antibodies
  2. Homo sapiens
  3. ZBTB11

ENCAB000BJN

Antibody against Homo sapiens ZBTB11

Homo sapiens
K562, GM12878, HepG2, MCF-7
characterized to standards with exemption
Status
released
Source (vendor)
Sigma
Product ID
SAB2102741
Lot ID
QC10553
Characterized targets
ZBTB11 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Antigen description
Peptide region of the protein sequence according to NP_055230.
External resources

Characterizations

ZBTB11 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
K562 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; SAB2102741). The IP fraction was loaded on a 12% Bio-Rad TGX gel and separated with the Bio-Rad Tetra Cell system. The whole lane was excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility. Analysis of whole lane 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, ZBTB11, was identified as the 3rd enriched protein and the 1st transcription factor based on IP-Mass Spectrometry.
Reviewer comment
Missing gel image
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 ZBTB11.pdf
ZBTB11 (Homo sapiens)
K562GM12878HepG2MCF-7
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Whole cell lysates of K562, HepG2, MCF7, and GM12878 were immunoprecipitated using the primary antibody (Sigma; SAB2102741). 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 Scientifiec). Protein Marker (PM) is labeled in kDa. No distinct bands around ~120 kDa were observed. Expected size ~119 kDa.
Submitter comment
--
Reviewer comment
Bands are not 50% of overall signal and they also appear in the control lanes. Rescued by mass spectrometry data
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
ZBTB11 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
An immunoprecipitation followed by Western blot was performed with Sigma SAB2102741 (lot QC10553) antibody against ZBTB11 in K562 whole cell lysate. The expected size of ZBTB11 is 119 kDa. No bands were detected. The experiment was repeated and the whole IP fraction was analyzed by mass spec in the secondary validation.
Submitter comment
non specific bands
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB