Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: Scientific Reports

Article Title: Tracing the invisible mutant ADNP protein in Helsmoortel-Van der Aa syndrome patients

doi: 10.1038/s41598-024-65608-x

Figure Lengend Snippet: Overview of the reported ADNP antibodies with indication of the observed molecular weights and tested sample materials.

Article Snippet: Adult human brain (NB820-59177), human brain cerebellum (NB820-59180), human brain frontal lobe (NB820- 59186), and human brain hippocampus whole tissue lysates (NB820-59187) from a healthy 45-year-old male subject were purchased from Novus Biologicals.

Techniques: Molecular Weight, Transfection, Expressing, Construct

Journal: Scientific Reports

Article Title: Tracing the invisible mutant ADNP protein in Helsmoortel-Van der Aa syndrome patients

doi: 10.1038/s41598-024-65608-x

Figure Lengend Snippet: A polyclonal N-terminal ADNP antibody from Aviva Systems detects ADNP specifically in murine and rat tissues and suggests proteolytic processing of the protein in the human brain. Cerebellum, frontal cortex or lobe, hippocampus and whole brains of control mice, rats and humans were lysed in RIPA buffer and used as protein samples for the assessment of N-terminal antibody of Aviva systems. (A – C) The predicted molecular weight of ADNP is 124 kDa. The antibody recognizes ADNP in a range of 145 kDa with (E) additional lower mass signal of 85 kDa in all human brain regions. (B – D – F) Western blot analysis of the blocking peptide competition assay. Supplementation of the immunization peptide in a 5 × excess to antibody concentration reduced the signal observed at 145 kDa in all tested cell lines. Importantly, the 85 kDa band suggestive for proteolytic cleavage as well as degraded ADNP signal disappeared completely after immunization peptide supplementation. GAPDH was used as a loading control.

Article Snippet: Adult human brain (NB820-59177), human brain cerebellum (NB820-59180), human brain frontal lobe (NB820- 59186), and human brain hippocampus whole tissue lysates (NB820-59187) from a healthy 45-year-old male subject were purchased from Novus Biologicals.

Techniques: Control, Molecular Weight, Western Blot, Blocking Assay, Competitive Binding Assay, Concentration Assay

Journal: Scientific Reports

Article Title: Tracing the invisible mutant ADNP protein in Helsmoortel-Van der Aa syndrome patients

doi: 10.1038/s41598-024-65608-x

Figure Lengend Snippet: Different C-terminal ADNP antibodies detect ADNP in the range of 150 kDa and suggest proteolytic processing of the protein in the brain. Cerebellum, frontal cortex or lobe, hippocampus and whole brains of control mice, rats, and humans were lysed in RIPA buffer and used as protein samples for the assessment with three C-terminal antibodies with the optimized dilutions listed in Table . GAPDH was used as a loading control. The predicted molecular weight of ADNP is 124 kDa. ( A ) C ) Murine samples indicate detection of ADNP in the range of 150 kDa with bands suggesting proteolytic processing at 50 kDa. ( D – F ) Rat samples indicate detection of ADNP in the range of 150 kDa with bands indicating proteolytic processing at 82 kDa after incubation with the C-terminal Abcam antibody. ( G – I ) Human brain samples indicate detection of ADNP at different molecular weights of 124 – 150 kDa in the adult frontal lobe and hippocampus and highlight the antibody differences in detection of ADNP. The three tested antibodies showed strong band signals at lower molecular weights, which could indicate proteolytic cleavage or degradation of the protein.

Article Snippet: Adult human brain (NB820-59177), human brain cerebellum (NB820-59180), human brain frontal lobe (NB820- 59186), and human brain hippocampus whole tissue lysates (NB820-59187) from a healthy 45-year-old male subject were purchased from Novus Biologicals.

Techniques: Control, Molecular Weight, Incubation

Journal: Nature Communications

Article Title: Mammalian brain glycoproteins exhibit diminished glycan complexity compared to other tissues

doi: 10.1038/s41467-021-27781-9

Figure Lengend Snippet: Protein lysate from a representative male mouse cortex and cerebellum with human plasma as a positive control was treated with or without PNGase F and visualized using biotinylated lectins (ConA, GNL, PHA-E, AAL, RCA, and SNA) in addition to immunoblotting for actin and staining for total protein. Non-specific binding of lectins to PNGase F is noted by an asterisk (*) near 35 kDa, as shown in the Total Protein stain. Protein blotting of brain lysate with each lectin has been repeated at least three times each with similar results. A schematic with common lectin-binding sites is shown for reference. Source data are provided as a Source Data file.

Article Snippet: Human Brain Cerebral Cortex Whole Tissue Lysate was purchased from Novus Biologicals (#NB820-59182), with 1mg used for glycomic analysis as described below.

Techniques: Clinical Proteomics, Positive Control, Western Blot, Staining, Binding Assay

Journal: Cell reports

Article Title: A PINK1 input threshold arises from positive feedback in the PINK1/Parkin mitophagy decision circuit

doi: 10.1016/j.celrep.2023.113260

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Healthy human brain lysate, whole (Novus Biologicals, NB820–59177).

Techniques: Recombinant, Bradford Assay, Mutagenesis, Plasmid Preparation, Microscopy, Cell Culture, Software, Sequencing

Journal: iScience

Article Title: Inhibition of the pseudokinase MLKL alters extracellular vesicle release and reduces tumor growth in glioblastoma

doi: 10.1016/j.isci.2022.105118

Figure Lengend Snippet:

Article Snippet: Human Brain Whole Tissue Lysate (Adult Whole Tumor) , Novus Biologicals , Cat# NB820-59423.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, EdU Assay, Flow Cytometry, Imaging, Cell Counting, Luciferase, Viability Assay, Isolation