1. Antibodies
  2. Homo sapiens
  3. ZBED1

ENCAB000AAK

Antibody against Homo sapiens ZBED1

Homo sapiens
K562, GM12878
characterized to standards
Status
released
Source (vendor)
Sigma
Product ID
WH0009189M1
Lot ID
11040-5G1
Characterized targets
ZBED1 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Isotype
IgG2aκ
Antigen description
ZBED1 (NP_004720, a.a. 3-100) partial recombinant protein with GST tag.
External resources

Characterizations

ZBED1 (Homo sapiens)
K562GM12878
Method: immunoblot
Attachment from submitter
Attachment from submitters
compliant
Caption
Whole cell lysates of K562 and GM12878 were immunoprecipitated using the primary antibody (Sigma; WH0009189M1). 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 ~82 kDa.
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB
ZBED1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not submitted for review by lab
Caption
K562 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0009189M1). 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
No IgG control and very faint band, but but is mostly supplementary to mass spec data
Reviewer comment
Barely visible band and no IgG. Rescued by mass spec
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
ZBED1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment from submitter
Attachment from submitters
compliant
Caption
Analysis of gel fragment 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, ZBED1, was identified as the 4th ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
Submitter comment
Rescues immunoblot and IP
Submitted by
Collin White
Lab
Richard Myers, HAIB