The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 1µg for 106 cells. ab91537-Mouse monoclonal IgG3, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
FunctionMajor role in the synthesis of nucleoside triphosphates other than ATP. Negatively regulates Rho activity by interacting with AKAP13/LBC. Acts as a transcriptional activator of the MYC gene; binds DNA non-specifically (PubMed:8392752). Exhibits histidine protein kinase activity.
Tissue specificityUbiquitously expressed.
Sequence similaritiesBelongs to the NDK family.
Cellular localizationCytoplasm. Nucleus. Isoform 2 is mainly cytoplasmic and isoform 1 and isoform 2 are excluded from the nucleolus.
IHC image of ab60602 staining in human cervical carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab60602, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Western blot - Anti-NME2 antibody (ab60602)
Anti-NME2 antibody (ab60602) at 1/1000 dilution + Recombinant Human NME2 protein (ab93692) at 0.1 µg
Secondary Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution developed using the ECL technique
Performed under reducing conditions.
Exposure time : 2 minutes
Flow Cytometry-Anti-NME2 antibody(ab60602)
Overlay histogram showing HeLa cells stained with ab60602 (red line). The cells were fixed with 80% methanol (5 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 (ab60602, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG3 [MG3-35] (ab18394, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
References for Anti-NME2 antibody (ab60602)
This product has been referenced in:
Yadav VK et al. Promoter-proximal transcription factor binding is transcriptionally active when coupled with nucleosome repositioning in immediate vicinity. Nucleic Acids Res42:9602-11 (2014).
Read more (PubMed: 25081206) »