mouse anti neurofilament 200 antibodies  (Boster Bio)

Journal: Materials Today Bio

Article Title: Hybrid construction of tissue-engineered nerve graft using skin derived precursors induced neurons and Schwann cells to enhance peripheral neuroregeneration

doi: 10.1016/j.mtbio.2024.101196

Figure Lengend Snippet: Schematic illustration of the present work and isolation, differentiation and characterization of SKPs, SKP-neurons and SKP-SCs. Schematic illustration (top) of a skin-derived precursor (SKP)-induced Schwann cell and SKP-induced neurons-mediated tissue-engineered nerve graft (TENG) for rat sciatic nerve repair. The consisting of chitosan nerve conduit, inserted silk fibroin fibers, SKP-neurons, SKP-SCs and their extracellular matrix can promote sciatic nerve regeneration and functional restoration nearly to the levels achieved by autologous nerve grafts according to comprehensive analysis including behavioral, electrophysiological, FG retrograde tracing, and histological evidence. (a) SKP spheres were generated from juvenile SKPs post 14 days of culture. (b–f) Characterizations by the immunostainings of fibronectin (green), nestin (red), sca-1(green), versican (red), and vimentin(green) for SKPs. (g) SKP spheres were induced to differentiate into SKP-neurons (h) and SKP-SCs (i) in vitro . (c) The characterization of SKP-neurons and SKP-SCs by immunostainings of NF200 (j1-j4), β tubulin III (k1-k4) for SKP-neurons and p75 NTR (l1-l4), S100β (m1-m4) for SKP-SCs. (red) neurons and SCs markers, (green) GFP, (blue) DAPI. Scale bar: 50 μm for a-f, 100 μm for g, and 20 μm for h-m.

Article Snippet: For immunofluorescent double-staining, rabbit anti-S100β polyclonal antibody (1:50 dilution) and mouse anti-neurofilament-200 (NF200) monoclonal antibody (1:200 dilution, both antibodies were from Sigma) were applied to nerve sections to allow incubation at 4 °C overnight, followed by further reaction with the secondary antibody (Goat anti-Mouse IgG-Alex-488, 1:500 and Donkey anti-Rabbit IgG-Cy3, 1: 1000) at 4 °C overnight, and nerve sections were observed under a confocal laser scanning microscope (TCS SP2, Leica).

Techniques: Isolation, Derivative Assay, Functional Assay, Retrograde Tracing, Generated, In Vitro

Journal: Materials Today Bio

Article Title: Hybrid construction of tissue-engineered nerve graft using skin derived precursors induced neurons and Schwann cells to enhance peripheral neuroregeneration

doi: 10.1016/j.mtbio.2024.101196

Figure Lengend Snippet: Construction of TENGs in vitro . (a–h) SEM images and (i–o) immunofluorescence images showed the micromorphology of GFP-SKP-neurons and GFP-SKP-SCs cultured on silk fibroin filaments and chitosan conduit 14 days post construction of TENGs. SCs markers (p75 NTR and S100 β), neurons markers (β tubulin III and NF200) and ECM molecules (collagen I, collagen IV, and laminin) were confirmed by immunostainings on the silk fibroin filaments.

Article Snippet: For immunofluorescent double-staining, rabbit anti-S100β polyclonal antibody (1:50 dilution) and mouse anti-neurofilament-200 (NF200) monoclonal antibody (1:200 dilution, both antibodies were from Sigma) were applied to nerve sections to allow incubation at 4 °C overnight, followed by further reaction with the secondary antibody (Goat anti-Mouse IgG-Alex-488, 1:500 and Donkey anti-Rabbit IgG-Cy3, 1: 1000) at 4 °C overnight, and nerve sections were observed under a confocal laser scanning microscope (TCS SP2, Leica).

Techniques: In Vitro, Immunofluorescence, Cell Culture

Journal: Materials Today Bio

Article Title: Hybrid construction of tissue-engineered nerve graft using skin derived precursors induced neurons and Schwann cells to enhance peripheral neuroregeneration

doi: 10.1016/j.mtbio.2024.101196

Figure Lengend Snippet: Evaluation of axonal regeneration on TENGs after nerve bridging surgery. (a–d) The immunofluorescence images of the longitudinal sections of TENGs at 4 dpi, 1, 2, 3 weeks post-surgery. (a’-d’) are the local magnified fields of (a–d). The dashed lines indicate regenerating axon terminals. (e) Histogram of the axon regeneration distances at the proximal stumps of TENGs are calculated (n = 8). The SKP-neurons and SKP-SCs are illustrated (GFP, green). The axons are NF200 positive, red. Scale bar, 1000 μm and 200 μm respectively. § p < 0.05 versus 4 d, * p < 0.05 versus 1 w, and # p < 0.05 versus 2 w. (f–l) The immunofluorescence images of the longitudinal sections of TENG, autograft, and scaffold groups at 1, 2 weeks post-surgery. The dashed lines indicate regenerating axon terminals. The NF200 positive axons are illustrated in red. (i, m) Histograms of the axon regeneration distances of TENG, autograft, and scaffold groups are calculated at 1, 2 weeks post-surgery (n = 8). * p < 0.05 versus scaffold group.

Article Snippet: For immunofluorescent double-staining, rabbit anti-S100β polyclonal antibody (1:50 dilution) and mouse anti-neurofilament-200 (NF200) monoclonal antibody (1:200 dilution, both antibodies were from Sigma) were applied to nerve sections to allow incubation at 4 °C overnight, followed by further reaction with the secondary antibody (Goat anti-Mouse IgG-Alex-488, 1:500 and Donkey anti-Rabbit IgG-Cy3, 1: 1000) at 4 °C overnight, and nerve sections were observed under a confocal laser scanning microscope (TCS SP2, Leica).

Techniques: Immunofluorescence

Journal: Materials Today Bio

Article Title: Hybrid construction of tissue-engineered nerve graft using skin derived precursors induced neurons and Schwann cells to enhance peripheral neuroregeneration

doi: 10.1016/j.mtbio.2024.101196

Figure Lengend Snippet: Analysis of regenerated axon density post-surgery. (a–e) The immunofluorescence images of the cross sections of regenerated nerve distal portion to TENG, autograft, scaffold groups and the contralateral uninjured side (normal). (a1-e1) The NF200 positive axons are illustrated in green. (a2-e2) The S100β Schwann cells or myelin structure are illustrated in red. Scale bar, 500 μm and 100 μm respectively. (f) Histogram of the regenerated axon density were shown and analyzed (n = 8). * p < 0.05 versus scaffold group and # p < 0.05 versus normal group.

Article Snippet: For immunofluorescent double-staining, rabbit anti-S100β polyclonal antibody (1:50 dilution) and mouse anti-neurofilament-200 (NF200) monoclonal antibody (1:200 dilution, both antibodies were from Sigma) were applied to nerve sections to allow incubation at 4 °C overnight, followed by further reaction with the secondary antibody (Goat anti-Mouse IgG-Alex-488, 1:500 and Donkey anti-Rabbit IgG-Cy3, 1: 1000) at 4 °C overnight, and nerve sections were observed under a confocal laser scanning microscope (TCS SP2, Leica).

Techniques: Immunofluorescence

Journal: Materials Today Bio

Article Title: Hybrid construction of tissue-engineered nerve graft using skin derived precursors induced neurons and Schwann cells to enhance peripheral neuroregeneration

doi: 10.1016/j.mtbio.2024.101196

Figure Lengend Snippet: Histological observation and morphometric analysis of the target muscles. (a, b) Histograms showing the wet weight ratio of total anterior tibialis and gastrocnemius muscle, and the cross-sectional area of gastrocnemius muscle fibers. Data are expressed as means ± SD. * p < 0.05 versus scaffold group, § p < 0.05 versus non-grafted group. (c, e, g, i, k) Masson's trichrome staining images are obtained at 12 weeks post-surgery, of the sectioned gastrocnemius muscle on the injured side in TENG (c), autograft (e), scaffold (g), non-grafted groups (i) and the contralateral uninjured side (k, normal), respectively. Scale bar, 50 mm. Photomicrographs (d, f, h, j, l and d’, f’, h’, j’, l’) of longitudinal sections of gastrocnemius muscles at the injured side after α-bungarotoxin staining of motor endplates (red) and NF200 (green) positive regenerated axons are obtained in three nerve grafting groups, nongrafted group and the contralateral uninjured side muscle at 12 weeks. Scale bar: 50 μm for low magnification images, and 20 μm for high magnification images. Histograms showing the cross-sectional area of gastrocnemius muscle fibers (m) and average percentage of collagen fiber area (n). Data are expressed as means ± SD. * p < 0.05 versus scaffold group, § p < 0.05 versus non-grafted group and # p < 0.05 versus normal group.

Article Snippet: For immunofluorescent double-staining, rabbit anti-S100β polyclonal antibody (1:50 dilution) and mouse anti-neurofilament-200 (NF200) monoclonal antibody (1:200 dilution, both antibodies were from Sigma) were applied to nerve sections to allow incubation at 4 °C overnight, followed by further reaction with the secondary antibody (Goat anti-Mouse IgG-Alex-488, 1:500 and Donkey anti-Rabbit IgG-Cy3, 1: 1000) at 4 °C overnight, and nerve sections were observed under a confocal laser scanning microscope (TCS SP2, Leica).

Techniques: Muscles, Staining