Immunofluorescent analysis of cortical neuron-glial cell culture from E20 rat stained with Anti-UCHL1 Antibody, at a dilution of 1:500, in red, and Anti-Vimentin Antibody (A85423 | 1:2,000, in green. The blue is DAPI staining of nuclear DNA. The Anti-UCHL1 Antibody produces strong staining of the cell body and dendrites in neurons. The Anti-Vimentin Antibody stains intermediate filaments in fibroblastic and developing glial cells.
Western blot analysis of equal amounts of different tissue and cell lysates using Anti-UCHL1 Antibody, at a dilution of 1:2,000, in green, and Anti-Actin Antibody (A85388 | 1:1,000, in red,: [Lane 1] protein standard, [Lane 2] rat brain, [Lane 3] mouse brain, [Lane 4] NIH-3T3, [Lane 5] HEK293, [Lane 6] HeLa and [Lane 7] SH-SY5Y cells. The single band at 24 kDa mark corresponds to UCHL1 protein which is detectable in CNS extracts and lysates of cells with neuronal properties but not in lysates of HeLa, NIH-3T3 and other non-neuronal cells. Actin is detected with apparent molecular weight of 42 kDa and provides an excellent loading control.
Rat mixed neuron/glial cultures stained with Anti-UCHL1 Antibody (green) and Anti-GFAP Antibody (A85419 | red). Blue is a DNA stain. Note that the Anti-UCHL1 Antibody stains neurons strongly and specifically, and that the staining is concentrated in the cell bodies, though some does extend into the dendrites also.
Blots of whole cell homogenate of the human SH-SY5Y neuroblastoma cell line stained with Anti-GAPDH Antibody (A85379 | Left Lane) and Anti-UCHL1 Antibody (Right Lane). The Anti-GAPDH Antibody stains a ~37kDa band while the Anti-UCHL1 Antibody stains a band at about 24kDa.
Western blot analysis of equal amount of total proteins from different cell and tissue lysates using Anti-GFAP Antibody [3E10] (A270551), at a dilution of 1:1,000, in green. The lanes contain: [Lane 1] protein standards (red), [Lane 2] primary rat cortical neuron-glial cells, [Lane 3] rat brain, [Lane 4] mouse brain, [Lane 5] cow cortex, [Lane 6] cow cerebellum, and [Lane 7] pig hippocampus. Bands around 50 kDa correspond to GFAP protein and its alternative transcripts and fragments. Anti-GFAP Antibody [3E10] (A270551) has stronger reactivity with cow GFAP and pig GFAP compared to rodent GFAP; and also reacts strongly with human native and recombinant GFAP (not shown). The same blot was probed with Anti-UCHL1 Antibody (A85349), a neuron cell marker, at a dilution of 1:2,000, in red. Anti-UCHL1 Antibody (A85349) reveals a 25 kDa band corresponding to UCHL1 protein detected in all preparations, acting as a positive loading control.
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