- PA5-18039 - Invitrogen Antibodies AIF1 antibody

Submitted references Lytic Cell Death in Specific Microglial Subsets Is Required for Preventing Atypical Behavior in Mice.

Chuang HC, Nichols EK, Rauch I, Chang WC, Lin PM, Misra R, Kitaoka M, Vance RE, Saijo K
eNeuro 2021 Jan-Feb;8(1)

Ketogenic Diet as a potential treatment for traumatic brain injury in mice.

Har-Even M, Rubovitch V, Ratliff WA, Richmond-Hacham B, Citron BA, Pick CG
Scientific reports 2021 Dec 7;11(1):23559

Microglial neuropilin-1 promotes oligodendrocyte expansion during development and remyelination by trans-activating platelet-derived growth factor receptor.

Sherafat A, Pfeiffer F, Reiss AM, Wood WM, Nishiyama A
Nature communications 2021 Apr 15;12(1):2265

The effect of rosmarinic acid on deformities occurring in brain tissue by craniectomy method. Histopathological evaluation of IBA-1 and GFAP expressions.

Özevren H, Deveci E, Tuncer MC
Acta cirurgica brasileira 2020;35(4):e202000406

Role of sex and high-fat diet in metabolic and hypothalamic disturbances in the 3xTg-AD mouse model of Alzheimer's disease.

Robison LS, Gannon OJ, Thomas MA, Salinero AE, Abi-Ghanem C, Poitelon Y, Belin S, Zuloaga KL
Journal of neuroinflammation 2020 Sep 29;17(1):285

Efficient control of Japanese encephalitis virus in the central nervous system of infected pigs occurs in the absence of a pronounced inflammatory immune response.

Redant V, Favoreel HW, Dallmeier K, Van Campe W, De Regge N
Journal of neuroinflammation 2020 Oct 23;17(1):315

High-Fat Diet-Induced Obesity Causes Sex-Specific Deficits in Adult Hippocampal Neurogenesis in Mice.

Robison LS, Albert NM, Camargo LA, Anderson BM, Salinero AE, Riccio DA, Abi-Ghanem C, Gannon OJ, Zuloaga KL
eNeuro 2020 Jan Feb;7(1)

Involvement of homodomain interacting protein kinase 2-c-Jun N-terminal kinase/c-Jun cascade in the long-term synaptic toxicity and cognition impairment induced by neonatal Sevoflurane exposure.

Liang L, Xie R, Lu R, Ma R, Wang X, Wang F, Liu B, Wu S, Wang Y, Zhang H
Journal of neurochemistry 2020 Aug;154(4):372-388

A Single Primary Blast-Induced Traumatic Brain Injury in a Rodent Model Causes Cell-Type Dependent Increase in Nicotinamide Adenine Dinucleotide Phosphate Oxidase Isoforms in Vulnerable Brain Regions.

Rama Rao KV, Iring S, Younger D, Kuriakose M, Skotak M, Alay E, Gupta RK, Chandra N
Journal of neurotrauma 2018 Sep 1;35(17):2077-2090

Nicotinic Acetylcholine Receptor α9 and α10 Subunits Are Expressed in the Brain of Mice.

Lykhmus O, Voytenko LP, Lips KS, Bergen I, Krasteva-Christ G, Vetter DE, Kummer W, Skok M
Frontiers in cellular neuroscience 2017;11:282

Ablation of CCAAT/Enhancer-Binding Protein Delta (C/EBPD): Increased Plaque Burden in a Murine Alzheimer's Disease Model.

Lutzenberger M, Burwinkel M, Riemer C, Bode V, Baier M
PloS one 2015;10(7):e0134228

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (PFA fixed) analysis of IBA1 using IBA1 Polyclonal Antibody (Product # PA5-18039) (6 µg/mL) in staining of paraffin embedded Human Spleen. Heat induced antigen retrieval with citrate buffer pH 6, HRP-staining.

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Experimental details: Negative Control for IBA1 Polyclonal Antibody (Product # PA5-18039), showing immunohistochemical staining of paraffin embedded Human Spleen, with no primary antibody.

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Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2a - IBA-1 immunostaining (Control group). IBA-1 positive expression was seen in microglia cells, blood vessel endothelial cells and nuclei of some neurons ( yellow arrow ) ( left parietal lobe ). Scale bar = 500 mum.

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Experimental details: Figure 2b - IBA-1 immunostaining (TBI group). Positive IBA-1 expression was observed in hyperplasia endothelial cells ( red arrow ), microglia cells and degenerative neurons in the blood vessel ( yellow arrow ) lumen ( left parietal lobe ). Scale bar = 500 mum.

Supportive validation

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Experimental details: Figure 2c - IBA-1 immunostaining (TBI+RA group). IBA-1 expression was found to be significant in endothelial cells in blood vessels ( red arrow ) and small microglia cells ( yellow arrow ) ( left parietal lobe ). Scale bar = 500 mum.

Supportive validation

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Experimental details: Fig. 4 Histopathology, lymphocyte infiltration, and microglial activation in the cerebrum. Representative histopathologic lesions like perineuronal cuffing ( a ), perivascular cuffing ( b ), and noduli of microglial cells ( c ) observed in the cerebrum of animals B16 and B17 are shown. Representative pictures of infiltrating CD3+ T cells as visualized in the cerebrum of animals D2 ( d ), B15 ( e ), and B16 ( f ) are shown. Quantification of the number of CD3+ T cells in the cerebrum was done by counting their presence in 4 regions of 0.0875 mm 2 under the fluorescence microscope of control pigs (D1, D2, D3, D8), pigs with a less than 5-fold increase in IFNy mRNA expression (A2, A3, A11, A13, A14, A15, A16, A17, A18, A22, A23, B2, B3, B11, B13, B14, B17, B22 B23), and pigs with more than 5-fold increase in IFNy expression (B15, B16). Each dot represents the mean of one animal ( g ). The activation status of microglial cells upon intradermal inoculation in the cerebrum was determined by quantification of IBA1 using the IHC profiler in ImageJ. Control pig D2 ( h ) shows a less intense IBA1 staining compared to pig B15 that showed an increased expression of pro-inflammatory cytokine mRNA ( i ). Even though scored as low positive, a more branched morphology of the microglial cells is seen in animal B16 as indicated by the arrows ( j )

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Experimental details: Figure 4. Numbers of neurons but not microglia are increased in the TRN in adult Casp1 -/- mice. This figure is supported by Extended Data Figure 4-1 . A , Enlarged image of the GFP + PI + cluster shown in Figure 3 . Mice were injected with PI as shown in Figure 3 A , and fetal brain slices were stained with anti-Pax6 antibody. Images of Cx3cr1 -GFP (top left), PI (top right), anti-Pax6 (bottom left), and merged signals are shown. Scale bar: 50 mum. B , Representative images of anti-NeuN staining signals from male Casp1 -/- ( N = 3, right, from 2 litters) and sex/age-matched WT ( N = 3, left, from 2 litters) adult brains are shown. TRN regions are outlined in yellow. Scale bar: 200 mum. C , Average anti-NeuN signals from Casp1 -/- (red triangles) and WT (black circles) mice are shown as cell number/mm 2 . Dots indicate individual animals and are the average cell numbers from a total of six serial sections stained as shown in B ; p = 0.0209, df = 4, t = 3.694. Error bars indicate SEM; * p < 0.05. D , Representative images of anti-Iba1 antibody staining in the TRNs (regions outlined in yellow) of male WT ( N = 3, left, from 2 litters) and Casp1 -/- ( N = 3, right, 2 litters) mice are shown. Scale bar: 200 mum. E , Average number of Iba1 + cells in Casp1 -/- (red triangles) and WT (black circles) mice are shown as mean cell number/mm 2 . Dots indicate individual animals and are the average of cell numbers from a total of six serial sections stained as shown in D ; p = 0.7533, df

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Experimental details: Figure 8 The double staining for alpha7, alpha9 or alpha10 nAChR subunits (red) and markers of microglia (Iba1, green, A,C,E ) or astrocytes glial fibrillary acidic protein (GFAP, green, B,D,F ) in the cerebellum (Crb), cortex (Crtx) or putramen (Put). Abbreviations: WM, white matter; GL, granular layer of cerebellum; V, internal pyramidal layer; VI, multiform layer of cortex. In (A) bar is 20 mum, in (B-F) 50 mum.

Supportive validation

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Experimental details: Figure 5 Microglia and astrocyte changes in the cortex and hippocampus of control (n = 5), KD (n = 5), mTBI (n = 5), and mTBI + KD (n = 5) mice. KD reduces the mTBI-induced elevation in active astrocyte expression in the DGH only and not in activated microglia expression at 30 days post-injury. Graphs present quantification of the total surface labeled with ( A ) GFAP in the DGH ( B ) GFAP in the temporal cortex ( C ) Iba-1 in the DGH ( D ) Iba-1 in the temporal cortex. In addition, ( E , F ) Astrocyte morphology in the DG Hilus region displayed approximately twice the ramification as the control samples. ( G ) Representative images of immunohistochemical staining in the DGH and the temporal cortex are presented. GFAP positive cells are shown in green, Iba-1 positive cells are shown in yellow, and NeuN+ cells in red. The scale bars are 20-25 mum.

Supportive validation

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Experimental details: Figure 6. A , B , Microglia density in the inner ( A ) and outer ( B ) dentate, as measured by the mean (+-SEM) percentage area covered by Iba1 + stain throughout the whole hippocampus and in the dorsal and ventral subregions. C , D , Phagocytic activity in the inner ( C ) and outer ( D ) dentate, as measured by the mean (+-SEM) percentage area covered by CD68 + stain throughout the whole hippocampus and in the dorsal and ventral subregions. E , F , Relative degree of microglial phagocytic activity in the inner ( E ) and outer ( F ) dentate, as measured by the mean (+-SEM) percentage of Iba1 + area also positive for CD68 throughout the whole hippocampus and in the dorsal and ventral subregions. G , H , Absolute microglial phagocytic activity in the inner ( G ) and outer ( H ) dentate, as measured by the mean (+-SEM) percentage area positive for colocalized Iba1 and CD68 throughout the whole hippocampus, and in the dorsal and ventral subregions. N = 10/group. * p < 0.05, ** p < 0.01, *** p < 0.001. I , Representative images of Iba1 + (green) and CD68 + (red) cells in the dorsal dentate gyrus. DAPI (blue) counterstaining was used to visualize nuclei, determine position relative to bregma, and create ROIs around the dentate gyrus. J , Representative image of inner and outer ROIs of the dentate gyrus. See Figure 6-1 for estimation statistics performed for microglia-related measures.

PA5-18039

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