ENCAB000BRJ

Antibody against Homo sapiens RNF2

Homo sapiens
K562, HepG2
characterized to standards
Status
released
Source (vendor)
Active Motif
Product ID
39664
Lot ID
23012002
Characterized targets
RNF2 (Homo sapiens)
Host
mouse
Clonality
monoclonal
Purification
Protein G
Isotype
IgG
Antigen description
antibody was raised against the recombinant full-length mouse protein.
External resources

Characterizations

RNF2 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment from submitter
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, RNF2, was identified as the 1st ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB
Download
RNF2 (Homo sapiens)
K562HepG2
Method: immunoprecipitation
Attachment from submitter
compliant
Caption
An immunoprecipitation followed by Western blot was performed with Active Motif 39664 (lot 23012002) antibody against RNF2 in K562 and HepG2 whole cell lysate along with a control IP with normal IgG from the same host species as the test antibody (mouse, Santa Cruz sc-2025). The expected size of RNF2 is 40 kDa. A band of ~40 kDa was detected in the test IP and not in the control IP. The experiment was repeated and this band was analyzed by mass spec in the secondary validation.
Submitted by
Flo Pauli-Behn
Lab
Richard Myers, HAIB
RNF2 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
not reviewed
Caption
Whole cell lysates of K562 and HepG2 were immunoprecipitated using the primary antibody (ActiveMotif; 39664). 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 ~40 kDa. Expected size ~37-38 kDa
Submitter comment
--
Reviewer comment
IP gel for mass spectrometry analysis
Submitted by
Mark Mackiewicz
Lab
Richard Myers, HAIB