Anti-c-Myc [9E11] antibody - ChIP Grade (ab56)
- Product nameAnti-c-Myc [9E11] antibody - ChIP GradeSee all c-Myc primary antibodies ...
- DescriptionMouse monoclonal [9E11] to c-Myc - ChIP Grade
- Conjugation notes-
- Tested applicationsFlow Cyt, ChIP, WB, IP, IHC-P, IHC-Fr more details
- Species reactivityReacts with: Chicken, Human, Saccharomyces cerevisiae
Synthetic peptide: AEEQKLISEEDL conjugated to KLH, corresponding to amino acids 408-420 of Human c-Myc.
- Positive control
- Purchase matching WB positive control:Active human c-Myc full length protein
- HL-60 cells or breast tumors. Flow Cyt: This antibody gave a positive signal in paraformaldehyde fixed/Tween permeabilised HL60 cells.
- General notes
This product is available conjugated to FITC see ab150238
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPhosphate buffered saline
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Monoclonal
- Clone number9E11
- Light chain typekappa
- Epigenetics and Nuclear Signaling
- Domain Families
- HLH / Leucine Zipper
- HLH / Leucine Zipper
Our Abpromise guarantee covers the use of ab56 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Flow Cyt: 1/200.|
|ChIP||ChIP: Use at an assay dependent concentration. 2ul per 500ug of extract, see H. Zhang et al.|
|WB||WB: 1/500 - 1/1000. Predicted molecular weight: 49 kDa. Additional non-specific bands observed at 75, 110, 140 kDa using mouse and human cells (see Abreview).|
|IP||IP: Use a concentration of 5 µg/ml.|
|IHC-P||IHC-P: Use at an assay dependent concentration.|
|IHC-Fr||IHC-Fr: Use at an assay dependent concentration.|
- FunctionParticipates in the regulation of gene transcription. Binds DNA in a non-specific manner, yet also specifically recognizes the core sequence 5'-CAC[GA]TG-3'. Seems to activate the transcription of growth-related genes.
- Involvement in diseaseNote=Overexpression of MYC is implicated in the etiology of a variety of hematopoietic tumors.
Note=A chromosomal aberration involving MYC may be a cause of a form of B-cell chronic lymphocytic leukemia. Translocation t(8;12)(q24;q22) with BTG1.
Defects in MYC are a cause of Burkitt lymphoma (BL) [MIM:113970]. A form of undifferentiated malignant lymphoma commonly manifested as a large osteolytic lesion in the jaw or as an abdominal mass. Note=Chromosomal aberrations involving MYC are usually found in Burkitt lymphoma. Translocations t(8;14), t(8;22) or t(2;8) which juxtapose MYC to one of the heavy or light chain immunoglobulin gene loci.
- Sequence similaritiesContains 1 basic helix-loop-helix (bHLH) domain.
modificationsPhosphorylated by PRKDC. Phosphorylation at Thr-58 and Ser-62 by GSK3 is required for ubiquitination and degradation by the proteasome.
Ubiquitinated by the SCF(FBXW7) complex when phosphorylated at Thr-58 and Ser-62, leading to its degradation by the proteasome. In the nucleoplasm, ubiquitination is counteracted by USP28, which interacts with isoform 1 of FBXW7 (FBW7alpha), leading to its deubiquitination and preventing degradation. In the nucleolus, however, ubiquitination is not counteracted by USP28, due to the lack of interaction between isoform 4 of FBXW7 (FBW7gamma) and USP28, explaining the selective MYC degradation in the nucleolus. Also polyubiquitinated by the DCX(TRUSS) complex.
- Cellular localizationNucleus > nucleoplasm. Nucleus > nucleolus.
- Formc-Myc is also expressed in the cytoplasm.
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Anti-c-Myc [9E11] antibody - ChIP Grade images
c-Myc was immunoprecipitated using 0.5mg CHO overexpressing Stra8 whole cell lysate, 5µg of Mouse monoclonal to c-Myc and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, CHO overexpressing Stra8 whole cell lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab56.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.
Band: 49KDa; c-Myc
Overlay histogram showing HL60 cells stained with ab56 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Anti-c-Myc [9E11] antibody - ChIP Grade (ab56) at 1/1000 dilution + whole cell lysate prepared from huh-7 cells at 15 µg
Anti-mouse IgG conjugated to HRP at 1/10000 dilution
Predicted band size : 49 kDa
Image courtesy of an anonymous Abreview.
References for Anti-c-Myc [9E11] antibody - ChIP Grade (ab56)
This product has been referenced in:
- Mirzaei H et al. Systematic measurement of transcription factor-DNA interactions by targeted mass spectrometry identifies candidate gene regulatory proteins. Proc Natl Acad Sci U S A 110:3645-50 (2013). ChIP . Read more (PubMed: 23388641) »
- Mittal AK et al. Role of CTLA4 in the proliferation and survival of chronic lymphocytic leukemia. PLoS One 8:e70352 (2013). WB ; Human . Read more (PubMed: 23936412) »