Recombinant full length human NSE purified from E. coli.
WB: Rat brain and spinal cord lysate; Mouse brain and spinal cord lysate; NIH/3T3, HEK-293, HeLa, SH-SY5Y and C6 cell lysate.
ICC/IF: Mixed cortical neuron-glial cell culture from E20 rat and HEP293 cells.
The total protein content in serum: about 60mg/ml.
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
1/500. For optimal signal, fix with 3.7% formalin in PBS for one minute followed by 100% cold (-20C) methanol for one minute.
Has neurotrophic and neuroprotective properties on a broad spectrum of central nervous system (CNS) neurons. Binds, in a calcium-dependent manner, to cultured neocortical neurons and promotes cell survival.
The alpha/alpha homodimer is expressed in embryo and in most adult tissues. The alpha/beta heterodimer and the beta/beta homodimer are found in striated muscle, and the alpha/gamma heterodimer and the gamma/gamma homodimer in neurons.
Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 4/5.
Belongs to the enolase family.
During ontogenesis, there is a transition from the alpha/alpha homodimer to the alpha/beta heterodimer in striated muscle cells, and to the alpha/gamma heterodimer in nerve cells.
Cytoplasm. Cell membrane. Can translocate to the plasma membrane in either the homodimeric (alpha/alpha) or heterodimeric (alpha/gamma) form.
All lanes : Anti-NSE antibody (ab64721) at 1/5000 dilution
Lane 1 : Protein standard Lane 2 : Rat brain lysate Lane 3 : Rat spinal cord lysate Lane 4 : Mouse brain lysate Lane 5 : Mouse spinal cord lysate Lane 6 : NIH/3T3 (mouse embryo fibroblast cell line) cell lysate Lane 7 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate Lane 8 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate Lane 9 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate Lane 10 : C6 (Rat glial tumor cell line) cell lysate
Immunofluorescence analysis of mixed cortical neuron-glial cell culture from E20 rat stained for NSE using ab64721 at a 1/500 dilution (red). Costained with chicken anti-GFAP at a 1/5000 dilution (green). The blue is Hoechst staining of nuclear DNA.
Immunocytochemistry/ Immunofluorescence of a mixed culture of cortical cells (4% PFA-fixation), with the red staining being GFAP, the green staining being NSE stained with ab64721, and the blue nuclear staining being DAPI.
Western blot - Anti-NSE antibody (ab64721)
Anti-NSE antibody (ab64721) + Adult male mouse brain at 10 mg/ml
Immunofluorescent staining of human embryonic kidney cell line 293 by ab64721 (1:500). Hek293 expresses many neuronal proteins.
The red channel shows staining with ab64721, which recognizes all of these 293 cells. The green channels shows staining for another neuronal marker with a monoclonal antibody to ubiquitin C-terminal hydrolase 1 (UCHL1), ab64722. This neuronal gene is apparently activated in a cell density dependent fashion and at this stage only a few cells express this protein. However all cells that express NSE also express UCHL1.