1. Antibodies
  2. Homo sapiens
  3. MITF

ENCAB000BSE

Antibody against Homo sapiens MITF

Homo sapiens
K562
characterized to standards with exemption
Homo sapiens
any cell type or tissue
partially characterized
Status
released
Source (vendor)
Active Motif
Product ID
39789
Lot ID
16210001
Characterized targets
MITF (Homo sapiens)
Host
mouse
Clonality
monoclonal
Purification
Protein G
Isotype
IgG
Antigen description
This MITF antibody was raised against an N-terminal fragment of human MITF
External resources

Characterizations

MITF (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, MITF, was identified as the 3rd ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
 MITF.png
MITF (Homo sapiens)
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not reviewed
Caption
K562 whole cell lysate was immunoprecipitated using the primary antibody (Active Motif; 39789). 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
MITF (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 (Active Motif; 39789). 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 ~78 kDa.
Submitter comment
The standards allow for using mass spec to rescue an immunoblot
Reviewer comment
Mass spec passes
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB
Download
 MITF_IP_WB.png