antibody specific for app biotinylated 22c11 clone  (Millipore)

Journal: Brain Pathology

Article Title: Detection and verification of neurodegeneration after traumatic brain injury in the mouse: Immunohistochemical staining for amyloid precursor protein

doi: 10.1111/bpa.13163

Figure Lengend Snippet: Immunofluorescent staining in naïve mice with the mouse monoclonal against amyloid precursor protein (APP), Clone 22C11. (A) Confocal photomicrographs acquired from a brain slice across the cortex and hippocampus in a Z‐stack of 10 μm. (B) Stained cortical pyramidal neurons and their apical dendrites ( triangles ) under higher magnification. Cytosolic staining pattern is highlighted in ( inset ). (C) and (D) No sharp somatic staining was observed in hippocampal pyramidal cells (pcl) and granule cells (gcl). Instead, high background staining was seen in dendritic areas within the hippocampus (ml, so, and sr). Perivascular staining indicated with ( double arrows ). (E) Astrocytic‐like staining in external capsule (ec), the major white matter bundle of our focus. (F) Intensely stained ependymal cells (ep) surrounding the lateral ventricles (lv). (G)–(I) Single confocal image showing double staining with 22C11 (green) and a rabbit monoclonal against glial fibrillary acidic protein (GFAP; red) in ec and hippocampus. Whereas GFAP staining could be seen all over the slice, 22C11‐stained fibrous structures were predominantly found in ec. After merge, 22C11 would stain the central part of GFAP‐verified astrocytes, resulting in partial colocaliztion (I, arrows ). Unlike ec, very small fraction of GFAP‐stained astrocytes in the hippocampus were costained with 22C11. hf, hippocampal fissure; so, stratum oriens; sr, stratum radiatum. Scale bars: 50 μm in (A); 10 μm in (B)–(I ) .

Article Snippet: Clone 22C11 (MAB 348, Millipore‐Sigma) is a mouse monoclonal antibody that was produced by immunizing mice with amino acids 66–81 of human APP at the N‐terminus (homologous with mouse N‐terminal peptide).

Techniques: Staining, Slice Preparation, Double Staining

Journal: Brain Pathology

Article Title: Detection and verification of neurodegeneration after traumatic brain injury in the mouse: Immunohistochemical staining for amyloid precursor protein

doi: 10.1111/bpa.13163

Figure Lengend Snippet: Validating specificity of 22C11 and Y188 by comparing staining of APP knockout and wild‐type mice. In wild‐type mice (A)‐(C), Y188 (green) showed sharp staining in hippocampal (A) and cortical neurons (B), and sparse staining in corpus callosum (cc, C). On the contrast, 22C11 (red) showed faint staining in these neurons. In addition to intense staining of ependymal cells, 22C11 also stained out astrocytes in cc. Arrowhead : perivascular staining. Double staining with both Y188 and 22C11 in the same cells and/or area is highlighted in insets . In APP null mice (D)–(F), we could not find green staining from Y188. However, 22C11 showed the same staining pattern as in wild‐type animals (A)–(C). As a result, double staining in APP knockouts exhibited 22C11 immunoreactivity (red) only. sl‐m, stratum lacunosum ‐ moleculare . Scale bars: 10 μm in all panels, including insets .

Article Snippet: Clone 22C11 (MAB 348, Millipore‐Sigma) is a mouse monoclonal antibody that was produced by immunizing mice with amino acids 66–81 of human APP at the N‐terminus (homologous with mouse N‐terminal peptide).

Techniques: Staining, Knock-Out, Double Staining

Journal: Brain Pathology

Article Title: Detection and verification of neurodegeneration after traumatic brain injury in the mouse: Immunohistochemical staining for amyloid precursor protein

doi: 10.1111/bpa.13163

Figure Lengend Snippet: Y188‐positive neuropathology could not be costained with 22C11. ( A ): Conical structures intensely stained by Y188 (green) 10 h after lFPI, still connecting to their basally stained parent cell bodies of cortical pyramidal neurons. (B): Most cell bodies could be stained with 22C11 (red), but not the axonal blebs, leaving cell bodies as orange to yellow and axonal blebs as pure green (C). (D)–(F): Discrete staining by Y188 of puncta and conical structures in ec from 22C11 staining in ependymal cells and astrocytes ( arrows ), without obvious co‐staining could be confirmed (F). (G)–(I): Positive staining of puncta and cones by CT‐20 (green), without co‐staining by 22C11 (red). Similarly, 22C11 (red) could not costain CT‐20‐stained axonal blebs (green) in the cortex ( inset ). Scale bars: 10 μm in all panels.

Article Snippet: Clone 22C11 (MAB 348, Millipore‐Sigma) is a mouse monoclonal antibody that was produced by immunizing mice with amino acids 66–81 of human APP at the N‐terminus (homologous with mouse N‐terminal peptide).

Techniques: Staining

Journal: Brain Pathology

Article Title: Detection and verification of neurodegeneration after traumatic brain injury in the mouse: Immunohistochemical staining for amyloid precursor protein

doi: 10.1111/bpa.13163

Figure Lengend Snippet: Immunoperoxidase (ABC) staining with 22C11 or Y188 10 hr after lFPI. (A)–(C) In naïve controls, 22C11 showed positive staining in the cortex and ec. (B) At higher magnification, some of cortical pyramidal neuron staining could be traced to the apical dendrites ( inset , arrow ). Some astrocytes also got stained (C, inset ), with a few punctate staining arranged in rows (triangles). (D)–(F) 10 h after lFPI, 22C11 showed a similar staining pattern in the cortex (E, inset , arrow ) and a largely increased number of astrocytes ( double arrows ) in ec (F, inset ). Meanwhile, some varicosity‐like rows got clearly stained in ec (F, inset, triangles ). Note that ependymal cells (ep) were intensely stained in both naïve (A) and (C) and injured mice (D) and (F). (G)–(I) In Naïve mice, Y188 clearly showed sharp staining of cortical neurons (H, inset ) and rows of oligodendrocytes in ec (I, inset , triangles ). (J)–(L) lFPI resulted in a high density of intensely stained puncta in ec and axonal blebs in cortex ( arrows ). At higher magnification, these conical axonal blebs possessed a swollen base (K, arrows ), with the apex pointing to basally stained cell bodies of cortical pyramidal neurons ( arrows , inset ). (L) In ec, a couple of conical structures equipped with a long tail ( arrows ) were evident among a large number of heavily stained puncta ( inset, arrow ). Likely due to the extremely high density in ec, it became difficult to differentiate rows of varicosity‐like staining after lFPI. Scale bars: 50 μm in (A), (D), (G), and (J); 20 μm in other panels; 10 μm in insets .

Article Snippet: Clone 22C11 (MAB 348, Millipore‐Sigma) is a mouse monoclonal antibody that was produced by immunizing mice with amino acids 66–81 of human APP at the N‐terminus (homologous with mouse N‐terminal peptide).

Techniques: Staining

Journal: Brain Pathology

Article Title: Detection and verification of neurodegeneration after traumatic brain injury in the mouse: Immunohistochemical staining for amyloid precursor protein

doi: 10.1111/bpa.13163

Figure Lengend Snippet: Summary diagram comparing two hypotheses on TBI consequences, suggested in different studies. (Left panel) By interpreting 22C11‐stained varicosities and spheroids persistent in WM as APP‐positive axonal swelling, previous studies on patient TBI have hypothesized that TBI‐induced disruption of axonal transport results in APP accumulation in axons, leading to axonal swelling (or damage) and neural dysfunction. (Right panel) Our data suggest an alternative hypothesis. Based on the observation of axonal blebs in gray matter and clusters of puncta in WM immediately after TBI, and condensed neuronal cell bodies and Wallerian degeneration at later stages, the present study suggests that TBI‐induced axonal truncation may initiate neuronal loss, and oligodendrocyte damage can cause demyelination and Wallerian degeneration of axons. Therefore, post‐TBI neural dysfunction may be a combined consequence of traumatic axonal truncation and oligodendrocyte damage. Stained pathologies in parentheses are interpreted as damaged structures in each hypothesis. Time window of observation, and staining method(s) for each pathology are also listed.

Article Snippet: Clone 22C11 (MAB 348, Millipore‐Sigma) is a mouse monoclonal antibody that was produced by immunizing mice with amino acids 66–81 of human APP at the N‐terminus (homologous with mouse N‐terminal peptide).

Techniques: Staining, Disruption