Human/Mouse/Rat MBP Antibody

MBP in Human Brain.

MBP was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Mouse Anti-Human/Mouse/Rat MBP Monoclonal Antibody (Catalog # MAB42282) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal processes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

MBP in Mouse Brain.

MBP was detected in perfusion fixed frozen sections of mouse brain using Mouse Anti-Human/Mouse/Rat MBP Monoclonal Antibody (Catalog # MAB42282) at 0.1 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to myelinated fibers. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

MBP in Rat Brain.

MBP was detected in perfusion fixed frozen sections of rat brain using Mouse Anti-Human/Mouse/Rat MBP Monoclonal Antibody (Catalog # MAB42282) at 0.1 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to myelinated fibers. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Detection of Human MBP by Immunohistochemistry

Pronounced myelin loss in FTD-GRN. (A, C) Representative western blots and (B, D) densitometric quantification of PLP, CNP, MBP, betaIII-tubulin ( betaIII-T) and neurofilament light chain (NF-L) in (A, B) superior frontal white matter, and (C, D) superior parietal white matter from control (n = 11), FTD-C9orf72 (n = 11), and FTD-GRN (n = 6) cases. Protein levels were normalised to beta-actin or GAPDH as a loading control, and are expressed relative to the mean of the control group. (E) Representative images and (F) myelination scores from LFB staining of superior frontal gyrus white matter from control (n = 5), FTD-C9orf72 (n = 4), and FTD-GRN (n = 3) cases from which tissue fixed for < 2 weeks was available. (G) Representative ASPA (red) and MBP (green) staining in superior frontal gyrus white matter, and (H) ASPA-positive cell density. Groups were compared by one-way ANOVA with Tukey’s post-test (B,D,H) or Kruskall-Wallis test with Dunn’s post-test (F): *p < 0.05; **p < 0.01; ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36967384), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human MBP by Immunohistochemistry

Pronounced myelin loss in FTD-GRN. (A, C) Representative western blots and (B, D) densitometric quantification of PLP, CNP, MBP, betaIII-tubulin ( betaIII-T) and neurofilament light chain (NF-L) in (A, B) superior frontal white matter, and (C, D) superior parietal white matter from control (n = 11), FTD-C9orf72 (n = 11), and FTD-GRN (n = 6) cases. Protein levels were normalised to beta-actin or GAPDH as a loading control, and are expressed relative to the mean of the control group. (E) Representative images and (F) myelination scores from LFB staining of superior frontal gyrus white matter from control (n = 5), FTD-C9orf72 (n = 4), and FTD-GRN (n = 3) cases from which tissue fixed for < 2 weeks was available. (G) Representative ASPA (red) and MBP (green) staining in superior frontal gyrus white matter, and (H) ASPA-positive cell density. Groups were compared by one-way ANOVA with Tukey’s post-test (B,D,H) or Kruskall-Wallis test with Dunn’s post-test (F): *p < 0.05; **p < 0.01; ***p < 0.001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36967384), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of MBP by Western Blot

GCN2 deficiency impacts the length of myelinated segments and myelin levels. Sections (30 μm) of the brain derived from two-month-old male WT (n = 3) and genetically deficient GCN2 (GCN2−/−, n = 3) mice were obtained and processed for immunofluorescence to visualize myelin segments. (A) Representative images showing the myelin basic protein (MBP) (red) staining used as a myelin marker, the nuclei DAPI (blue), and the composed image (merge). An augmented section (white box) of the images are shown at the right (Zoom). Arrows on the zoom image show the type of MBP-positive objects quantified. (B) Images obtained in A were quantified using the Image J software (1.53t version). The plot shows the number and length of myelin segments (upper panel) and the average length (lower panel) of myelin segments found in WT and GCN2−/− mice. (C) Protein extracts were prepared from brains derived from two-month-old WT (n = 4) and GCN2−/− (n = 4) mice. MBP levels were analyzed by Western blot using alpha-Tubulin as a loading control (left). Each lane represents an individual (30 μg/lane). Densitometry analysis of the blot was performed by ImageJ to quantify the intensities of the two MBP bands and then normalized to alpha-Tubulin (right) levels. Scale bar: 100 μm. **** p < 0.00001, * p < 0.05. t-test analysis. Error bars represent the mean ± SEM. The whole Western blot membrane is shown in Supplementary Figure S1. The MBP quantification strategy is described in the “Materials and Methods” section. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40004088), licensed under a CC-BY license. Not internally tested by R&D Systems.