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)
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
- Grant
- U54HG006998
- Download
- RNF2-K.png
RNF2 (Homo sapiens)
K562HepG2
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
- Grant
- U54HG006998
- Download
- RNF2_39664_23012002_IP-Western.png
RNF2 (Homo sapiens)
K562
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
- Grant
- U54HG006998
- Download
- RNF2_IP-Commassie1.png