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1) Product Images from "Accumulation of Misfolded SOD1 in Dorsal Root Ganglion Degenerating Proprioceptive Sensory Neurons of Transgenic Mice with Amyotrophic Lateral Sclerosis"
Article Title: Accumulation of Misfolded SOD1 in Dorsal Root Ganglion Degenerating Proprioceptive Sensory Neurons of Transgenic Mice with Amyotrophic Lateral Sclerosis
Journal: BioMed Research International
Figure Legend Snippet: ((a)–(c)) Misfolded SOD1 (misSOD1) immunolabeling using AJ10 antibody (red) reveals highly immunoreactive neuronal somata within the DRG (delimited in (a)) of end-stage (P120) SOD1 G93A mice. Some neuronal somata display cytoplasmic fragmentation indicative of degenerative changes (arrows in (b)); nuclei, counterstained with DAPI (blue), do not show apoptotic morphology (arrows in (c)). ((d)–(g)) Misfolded SOD1 immunolabeling (red) was combined with an anti-CD68 antibody (green) to reveal activated macrophagic cells; DAPI (blue) was used for nuclear staining. A degenerating neuron expressing misfolded SOD1 (delimited in (d)) and displaying clustered profiles of CD68 positive phagocytic cells is shown in high magnification in (e)–(g). (h) A frequency distribution profile of the size of the DRG neuron somata containing misfolded SOD1 with respect to that of the whole neuronal population; note that misfolded SOD1 positive neurons belong to the large-sized (presumably proprioceptive) population. Scale bars: 80 μ m in (a), 40 μ m in (b), (c), (d), and (g) (also valid for (e) and (f)).
Techniques Used: Immunolabeling, Mouse Assay, Staining, Expressing
Figure Legend Snippet: Micrographs taken from the DRGs of end-stage (P120) SOD1 G93A mice. Misfolded SOD1 (misSOD1) immunolabeling using AJ10 antibody (red) was combined with several markers for DRG neuronal types, as indicated (green). ((a)–(i)) Misfolded SOD1 immunoreactive neurons do not colocalize with the neuropeptides CGRP and SP, or with IB4, all of which are markers for neuronal types other than proprioceptive. ((j)–(l)) However, neurons containing misfolded SOD1 colocalize with PV, a marker for proprioceptive sensory neurons. In some cases, DAPI was used for nuclear staining (blue). The prominent background observed in (c) and (d) is due to an unspecific connective tissue reaction involving the secondary antibody when a mouse monoclonal was used as a primary antibody. Scale bars = 200 μ m.
Techniques Used: Mouse Assay, Immunolabeling, Marker, Staining
Figure Legend Snippet: ((a)–(d)) Double immunolabeling with TUJ1 (to label neuronal profiles, red), and anti-CD68 (to reveal microglial cells, green) antibodies in ventral ((a), (c)) and dorsal ((b), (d)) horn of WT ((a), (b)) and end-stage SOD1G93A ((c), (d)) mice; L: lateral, M: medial. CD68-positive profiles were rarely detectable in WT samples, but they were present in either large or moderate quantities in the ventral and dorsal horn, respectively, of SOD1 G93A samples. In SOD1 G93A dorsal horns, it was possible to observe some infiltrating microglial cells adjacent to medially located (presumably proprioceptive) axon fascicles, entering the spinal cord (arrows in (d)). (e) Quantification of CD68-positive profiles in the spinal cord of WT and end-stage SOD1 G93A mice. Bars represent mean ± SEM values for counts performed in 6–11 fields of 2 animals per experimental condition; *** P
Techniques Used: Immunolabeling, Mouse Assay
Figure Legend Snippet: The ultrastructural morphology of VRs ((a), (c), and (d)) and DRs (b) from end-stage SOD1 G93A mice. From a qualitative point of view, similar Wallerian-like degenerative changes could be observed in both VRs (a) and DRs (b); phagocytic cells (shaded in green), in which the nucleus is dashed in blue, appear to engulf the myelin debris (shaded in red); apparently normal myelinated axons are dashed in yellow and Schwann cell profiles are dashed in magenta. ((c)–(d)) In ventral roots it is possible to observe some regenerating axons (yellow in (c)) growing inside the ghosts of folded basal lamina profiles (green in (c)) filled with Schwann cell pedicles (magenta in (c)). Occasionally, a large growth cone (yellow in (d)) filled with organelles and surrounded by Schwann cell profiles (magenta in (d)) can be detected. Scale bars: 5 μ m in (a), 2.5 μ m in (b), 0.5 μ m in (c), and 1 μ m in (d).
Techniques Used: Mouse Assay
Figure Legend Snippet: Morphometric analysis of motor (L4 VRs, (a)–(f)) and sensory (L4 DRs, (g)–(l)) axons from WT and SOD1 G93A mice. An analysis was performed on 1 μ m semithin plastic transversal sections ((b), (c), (h), and (i)). ((a)–(c) and (g)–(i)) Counts showed a significant loss of the total number of motor, but not sensory, axons in the end-stage (P120) SOD1 G93A mice ((a) and (g)). ((d)–(f) and (j)–(l)) A frequency distribution analysis of myelinated axon size in WT VRs showed a clear bimodal profile (f) indicative of axons coming from α -MNs (large) and γ -MNs (small); note the selective loss of large axons in SOD1 G93A mice. Representative images of WT and SOD1 G93A ventral nerve are shown in (d) and (e); note the presence of abundant degenerating axons in SOD1 G93A animals. Although there was no evidence of the loss of sensory axons in dorsal nerve roots in SOD1 G93A mice, a more detailed examination of nerve profile morphology ((j) and (k)) revealed the presence of moderate numbers of axons exhibiting significant degrees of swelling (∗) and other degenerating features. A frequency distribution analysis of myelinated axon size in dorsal nerve roots (l) reflected the appearance of a new population of large diameter (degenerating) sensory axons in SOD1 G93A samples (indicated by a double arrowed line). The bars in the graphs represent the mean ± SEM values of counts performed in 2–11 animals per age and experimental condition; ** P
Techniques Used: Mouse Assay
Figure Legend Snippet: (a) Semithin plastic section of a DRG from end-stage (P120) SOD1 G93A mice showing a degenerating, microvacuolated, large neuronal soma, which is surrounded by apparently normal medium-sized neuronal cells. ((b) and (c)) The cells indicated in the semithin sections were examined by electron microscopy which revealed a well-preserved organelle structure in the medium-sized neuronal cell (b) and extensive microvacuolization in the degenerating large soma (c). (d) A large myelinated axonal profile adjacent to the DRG sensory neurons shows an accumulation of highly vacuolated mitochondria with a comparable morphology to that of degenerating mitochondria typically described in motor axons of SOD1 G93A mice (as shown in (e), taken from a sample of facial nucleus). Scale bars: 20 μ m in (a), 0.5 μ m in (b) and (c), and 1.5 μ m in (d) and (e).
Techniques Used: Mouse Assay, Electron Microscopy