ENCAB000ATW
Antibody against Homo sapiens ZHX2
Homo sapiens
K562
characterized to standards
Homo sapiens
HepG2, GM12878, MCF-7
characterized to standards with exemption
- Status
- released
- Source (vendor)
- GeneTex
- Product ID
- GTX112232
- Lot ID
- 40107
- Characterized targets
- ZHX2 (Homo sapiens)
- Host
- rabbit
- Clonality
- polyclonal
- Isotype
- IgG
- Antigen description
- Recombinant fgmt within amino acids 533 and 826 of ZHX2 (Uniprot ID#Q9Y6X8)
- External resources
Characterizations
ZHX2 (Homo sapiens)
compliant
- Caption
- Analysis of gel fragment 1 from HepG2: 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, ZHX2, was identified as the 1st ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
- Reviewer comment
- Rescues failed immunoblot.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- ZHX2-1_mass spec.png
ZHX2 (Homo sapiens)
compliant
- Caption
- Analysis of gel fragment 2 from HepG2: 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, ZHX2, was identified as the 1st ranked enriched protein and the 1st ranked transcription factor based on IP-Mass Spectrometry.
- Reviewer comment
- Rescues failed immunoblot.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- ZHX2-2_mass spec 2.png
ZHX2 (Homo sapiens)
HepG2
not reviewed
- Caption
- HepG2 whole cell lysate was immunoprecipitated using the primary antibody (GeneTex; GTX112232). The IP fraction was loaded on a 12% Bio-Rad TGX gel and separated with the Bio-Rad Tetra Cell system. Gel fragments (rectangle outline) corresponding to the bands indicated on the Coomassie Blue stained gel image were excised and sent to the University of Alabama at Birmingham Cancer Center Mass Spectrometry/Proteomics Shared Facility.
- Submitter comment
- None
- Reviewer comment
- Missing IgG Control. Submitted as gel for the secondary mass spec characterization.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- ZHX2_IP-MS-WB.png
ZHX2 (Homo sapiens)
GM12878HepG2MCF-7
exempt from standards
- Caption
- Whole cell lysates of GM12878, HepG2, and MCF7 were immunoprecipitated using the primary antibody (GeneTex; GTX112232). 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. Two bands were detected at ~53 and ~117kDa.
- Submitter comment
- --
- Reviewer comment
- Band of interest is not 50% of overall signal in lane, is a doublet, and in some cases, appears slightly in control lane. Rescued by mass spec.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- ZHX2_IP-WB.png
ZHX2 (Homo sapiens)
K562HepG2
compliant
- Caption
- Whole cell lysates of K562 and HepG2 were immunoprecipitated using the primary antibody (GeneTex; GTX112232). 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. Two bands were detected at ~43 and ~100kDa.
- Submitter comment
- None
- Reviewer comment
- Hep G2 band not 50% of signal, but rescued by mass spec. GM12878 is compliant
- Submitted by
- Flo Pauli-Behn
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- ZXH2_IP-WB.png