ENCAB000BHV
Antibody against Homo sapiens MYNN
Homo sapiens
K562, GM12878
characterized to standards with exemption
- Status
- released
- Source (vendor)
- Sigma
- Product ID
- WH0055892M1
- Lot ID
- 06132-3E8
- Characterized targets
- MYNN (Homo sapiens)
- Host
- mouse
- Clonality
- monoclonal
- Isotype
- IgG2aκ
- Antigen description
- MYNN (NP_061127, a.a. 501-611) partial recomb protein w/ GST tag. MW of GST tag is 26 kDa.
- External resources
Characterizations
MYNN (Homo sapiens)
compliant
- Caption
- K562 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0055892M1). 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. 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, MYNN, was identified as the 5th enriched protein and the 1st transcription factor based on IP-Mass Spectrometry.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- MYNN-1.png
MYNN (Homo sapiens)
compliant
- Caption
- K562 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0055892M1). 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. Analysis of gel fragment 2 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, MYNN, was identified as the 2nd enriched protein and the 1st transcription factor based on IP-Mass Spectrometry.
- Submitted by
- Aditi Narayanan
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- MYNN-2.png
MYNN (Homo sapiens)
not compliant
- Caption
- K562 whole cell lysate was immunoprecipitated using the primary antibody (Sigma; WH0055892M1). 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. Analysis of gel fragment 3 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, MYNN, was not identified based on IP-Mass Spectrometry.
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- MYNN-3.png
MYNN (Homo sapiens)
K562
not submitted for review by lab
- Submitted by
- Mark Mackiewicz
- Lab
- Richard Myers, HAIB
- Grant
- U54HG004576
- Download
- MYNN_IP-Commassie1.png
MYNN (Homo sapiens)
K562GM12878
exempt from standards
- Caption
- Whole cell lysates of K562 and GM12878 were immunoprecipitated using the primary antibody (Sigma; WH0055892M1). 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. Three bands were detected at ~42, 65, and 130 kDa. Expected size ~68-69 kDa
- Submitter comment
- --
- Reviewer comment
- Non-specific bands and no IgG control, but rescued by mass spec analysis
- Submitted by
- Flo Pauli-Behn
- Lab
- Richard Myers, HAIB
- Grant
- U54HG006998
- Download
- MYNN_IP-WB.png