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.
Flow Cytometry - Anti-NSE antibody (ab53025)This image is taken from an anonymous abreview.
Intracellular flow analysis of NSE on N2A mouse neuroblastoma cells using ab53025 at 1:500 dilution. Purple histogram represents rabbit IgG negative control; green line represents anti-NSE antibody (ab53025).
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-NSE antibody (ab53025)This image is courtesy of an Abreview submitted by Olga Cravetchi
ab53025 staining NSE in Rat brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.3 Triton-X, and blocked with 5% serum for 1 hour at room temperature. The sample was incubated with primary antibody (1/250) at 4°C for 16 hours. Ab96921, DyLight® 594-conjugated Donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
ICC/IF image of ab53025 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab53025, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemical analysis of paraffin-embedded
human brain tissue using ab53025 at 1/50-1/100 dilution. Left: without immunizing peptide; Right: with immunizing peptide
Western blot - Anti-NSE antibody (ab53025)
All lanes : Anti-NSE antibody (ab53025) at 1/500 dilution
Lane 1 : Extracts from HepG2 cells with no immunizing peptide Lane 2 : Extracts from HepG2 cells with immunizing peptide
Predicted band size : 47 kDa Observed band size : 47 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NSE antibody (ab53025)Image from Yu Yang. et. al. Mol. Cell. Proteomics, Mar 2009 (Fig 5).
ab53025 staining NSE in mouse brain tissue section by Immunohistochemistry (PFA-perfussion fixed frozen tissue sections). Tissues were collected from mice in each group at 10 weeks. Mice were euthanized, the brains were exposed and removed from the body. Then the brain was cut, on ice and fixed in 4% polyformaldehyde or glutaraldehyde solution for histological examination. The primary antibody was used at 1/75 dilution in PBS and incubated with sample for 2 hours at 37°C. A HRP-conjugated secondary antibody was used at for 1hr at 37°C in a humidity box.
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