3-diaminobenzidine Search Results


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  • 99
    Vector Laboratories dab peroxidase hrp substrate kit with nickel 3 3 diaminobenzidine
    Distribution of <t>DAB</t> CR + and DAB CB + cells is not the same across all RE internal subdivisions A: Brightfield images showing the distribution of DAB CR + cells in RE across the rostro-caudal axis of the thalamus. CR + cell area density varied depending on the subdivision of RE in which they were located. B: Distribution of DAB CB + cells. When compared, CR + and CB + cell distribution in all RE’s subregions and across the rostral to caudal levels did not appear to be the same. Overlay shown adapted from Swanson (2018) to highlight all RE internal subdivisions. Scale bar = 100μm. C: Comparison of DAB CR + and DAB CB + cell area density (cells/0.01mm 2 ) in all subdivisions of RE across the rostro-caudal axis. CB + cell densities were higher than CR + cell densities (except REm). Additionally, REl, REv and REcd subdivisions exhibited large CB + cell densities compared to other RE subregions. A moderate size effect was found for CB + cell area density across all levels and RE subdivisions (Hedges’ d=0.32). Abbreviations: β, bregma; CB, calbindin; CR, calretinin; DAB, <t>3,3’-Diaminobenzidine;</t> PRe, perireuniens, RE, nucleus reuniens of the thalamus, REa, reuniens rostral division anterior part; REd, reuniens rostral division dorsal part; REl, reuniens rostral division lateral part; REm, reuniens rostral division median part; REv, reuniens rostral division ventral part; REcm, reuniens caudal division median part; REcd, reuniens caudal division dorsal part; REcp, reuniens caudal division posterior part.
    Dab Peroxidase Hrp Substrate Kit With Nickel 3 3 Diaminobenzidine, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 492 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 3 3 diaminobenzidine
    Immunohistochemical staining of claudin-7 expression in (A and B) normal endometrium and (C and D) cancer tissues. Original magnification, (A and C) ×200 and (B and D) ×400; staining, DAB. (E) Claudin-7 mRNA expression in endometrial cancer cell lines, as detected by real-time reverse trancription-polymerase chain reaction analysis. N, normal tissues; T, tumor tissues; DAB, <t>3,3′-diaminobenzidine.</t>
    3 3 Diaminobenzidine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1453 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore 3 3 diaminobenzidine
    Avulsion of dorsal roots dramatically disrupted the dorsal spinal cord, causing an influx of blood cells only at the lesion site, while rhizotomy did not result in blood cell influx. Incubation of tissue with <t>3′,3-diaminobenzidine</t> (DAB) revealed
    3 3 Diaminobenzidine, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 17403 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies 3 3 diaminobenzidine
    Representative images of GFAP+ astrocytes in the hippocampus of adult Long-Evans hooded male rats exposed to low-F − chow/RO-H 2 O or low-F − chow/20 ppm F − drinking water beginning on gestational day 6. ( a ) Suprapyramidal blade of the dentate gyrus. ( b ) CA1 pyramidal cell layer. Cells displayed normal process-bearing morphology with no evidence of hypertrophy. <t>3,3-diaminobenzidine</t> staining (brown). Hematoxylin counterstain (blue) showed no disruption of the normal morphology of the hippocampal regions and no evidence of neuronal death. ( n = 6). Scale bar = 100 μm
    3 3 Diaminobenzidine, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 95/100, based on 8814 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore 3 3 diaminobenzidine tetrahydrochloride
    Detection of ROS and expression of ROS marker genes in inbreds with contrasting levels of submergence tolerance. (a.) Detection of H 2 O 2 and superoxide in selected submergence tolerant and sensitive maize genotypes. A subset of four nested association mapping panel parents were selected based on visual leaf scoring for ROS staining assays after submergence. Hydrogen peroxide was detected using stain <t>3,3′-diaminobenzidine</t> <t>tetrahydrochloride</t> (DAB), while superoxide was detected using nitroblue tetrazolium (NBT). Both assays were performed after 96 h of submergence. In B97, the third leaf was barely emerged and not used for the assay. (b-d) Real-time quantitative PCR of ROS genes. ROS genes were selected based on the study by Gadjev, Vanderauwera and Gechev [ 30 ] and showed a significant induction in response to at least two stresses that induce ROS formation in Arabidopsis. (b.) ALTERNATIVE OXIDASE 1a ( AOX1a ); (c.) WRKY6 (d.) CYTOCHROME P81 D8 ( CYP81D8 ). All sample were collected at 24 h and 72 h after submergence. Expression levels are relative to shoot tissue of control plants at 24 h. Letters above bars indicate nonsignificant differences in expression determined using Tukey’s HSD ( p
    3 3 Diaminobenzidine Tetrahydrochloride, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7492 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies 3 3 diaminobenzidine tetrahydrochloride
    Immunohistochemical analysis by MsMab-1 against tissue microarray of osteosarcomas. Osteosarcoma tissue microarray was stained with MsMab-1 followed by LSAB kit. Color was developed using DAB, and was counterstained with hematoxylin. Typical results were shown: (A) No. 3, (B) No. 5, (C) No. 13, (D) No. 15, (E) No. 18, (F) No. 24, (G) No. 26, (H) No. 31, and (I) No. 32. Insets show that MsMab-1 stained cytoplasm (A–I). Magnification: 200×. MsMab, multi-specific anti-mutated IDH1/2 mAb; DAB, <t>3,3-diaminobenzidine</t> <t>tetrahydrochloride.</t>
    3 3 Diaminobenzidine Tetrahydrochloride, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 2721 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies 3 3 diaminobenzidine dab
    Representative images (20x magnification) of airway sections stained for cathepsins. Immunostaining of cathepsins and corresponding isotype controls for cathepsins D (A–D), H (E–H) and K (I–L) from non-diseased and asthmatic sections. Specific staining was detected using a chemical chromophore <t>DAB</t> (brown) and cell nucleus was counterstained with haematoxylin (blue). Abbreviations DAB = <t>3,3′-diaminobenzidine.</t>
    3 3 Diaminobenzidine Dab, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 1470 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore 3 3 diaminobenzidine tetrahydrochloride dab
    Sections from control mice that underwent direct staining (i.e., no deparaffinization) revealed staining of nuclei (and sometimes distinct cytoplasmic staining) of neurons, myelin, and neuropil staining. (a and b) Staining in the cortex is revealed in nuclei, cytoplasm, and neuropil. (c) The hippocampal CA1 neurons are densely stained. Splotches of staining are apparent in the cortex and hippocampus. (d) Myelin staining in the globus pallidus (right) and caudate or putamen (left) is apparent among staining of cells (dark dots) and neuropil across the field. Some splotches of staining are also apparent (lower left). (e) Dark staining of myelin and neuropil is present in the substantia nigra. Staining of cells (dark dots) across the section is also apparent. (f) Eliminating the potassium ferrocyanide from the staining solution reveals an absence of labeling. Hippocampus is shown (compare to c). Note, some tissue artifact (upper right) is lightly labeled showing that the section was exposed to the <t>DAB</t> solution, and an extremely light depiction of the CA1 neurons can be made out. (g) Staining of the cerebellum reveals staining of granule cells, Purkinje neurons, and staining of neuropil and neurons in the molecular layer, which also has a heterogeneous pattern of staining. Bar in a = 20 µ (a,b). Bar in c = 100 µ (c–g). Note . DAB = <t>3,3′-diaminobenzidine</t> <t>tetrahydrochloride.</t>
    3 3 Diaminobenzidine Tetrahydrochloride Dab, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 906 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies 3 3 diaminobenzidine chromogen
    Effects of midazolam on ethanol-induced neuroapoptosis in mice. (A) Immunohistochemical analysis of brain sections (n=5/group) was performed using anti-C-3A and was visualized using <t>3,3′-diaminobenzidine</t> chromogen. The C-3A positive staining was documented under a light microscope and scoring of C-3A-positive cells per mm 2 brain sections was performed. The total number of C-3A-positive cells was presented as the percentage of positive cells compared to the control group. (B) TUNEL staining of brain sections (n=5/group) was performed and analyzed. Data are presented as the mean ± standard deviation. *P
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    Dojindo Labs 3 3 diaminobenzidine
    In situ hybridization of Notch2 and immunohistochemistry of SOX2 in a sagittal section of rat pituitary gland at embryonic day 14.5. a ) Hematoxylin and eosin staining. b ) In situ hybridization of Notch2 . c ) Negative control of in situ hybridization using a sense probe of Notch2 . d ) Double staining of Notch2 detected by in situ hybridization and SOX2 detected by immunohistochemistry. Immunoreactivity for SOX2 was revealed by <t>3,3'-diaminobenzidine</t> (brown) and the Notch2 in situ hybridization signal with nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate (blue). Arrow: prospective pars intermedia , Double arrow: prospective pars distalis , Triple arrow: prospective pars tuberalis . Bar = 100 μm.
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    Millipore 3 3 diaminobenzidine tablets
    In situ hybridization of Notch2 and immunohistochemistry of SOX2 in a sagittal section of rat pituitary gland at embryonic day 14.5. a ) Hematoxylin and eosin staining. b ) In situ hybridization of Notch2 . c ) Negative control of in situ hybridization using a sense probe of Notch2 . d ) Double staining of Notch2 detected by in situ hybridization and SOX2 detected by immunohistochemistry. Immunoreactivity for SOX2 was revealed by <t>3,3'-diaminobenzidine</t> (brown) and the Notch2 in situ hybridization signal with nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate (blue). Arrow: prospective pars intermedia , Double arrow: prospective pars distalis , Triple arrow: prospective pars tuberalis . Bar = 100 μm.
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    Zhongshan Golden Bridge Company 3 3 diaminobenzidine
    Immunohistochemistry staining for Notch1- and Notch3-positive cells in lung adenocarcinoma tissues of smokers and non-smokers, as determined using <t>3,3′-diaminobenzidine</t> and hematoxylin staining (magnification, x200). (A) Notch1-positive cells
    3 3 Diaminobenzidine, supplied by Zhongshan Golden Bridge Company, used in various techniques. Bioz Stars score: 92/100, based on 575 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Distribution of DAB CR + and DAB CB + cells is not the same across all RE internal subdivisions A: Brightfield images showing the distribution of DAB CR + cells in RE across the rostro-caudal axis of the thalamus. CR + cell area density varied depending on the subdivision of RE in which they were located. B: Distribution of DAB CB + cells. When compared, CR + and CB + cell distribution in all RE’s subregions and across the rostral to caudal levels did not appear to be the same. Overlay shown adapted from Swanson (2018) to highlight all RE internal subdivisions. Scale bar = 100μm. C: Comparison of DAB CR + and DAB CB + cell area density (cells/0.01mm 2 ) in all subdivisions of RE across the rostro-caudal axis. CB + cell densities were higher than CR + cell densities (except REm). Additionally, REl, REv and REcd subdivisions exhibited large CB + cell densities compared to other RE subregions. A moderate size effect was found for CB + cell area density across all levels and RE subdivisions (Hedges’ d=0.32). Abbreviations: β, bregma; CB, calbindin; CR, calretinin; DAB, 3,3’-Diaminobenzidine; PRe, perireuniens, RE, nucleus reuniens of the thalamus, REa, reuniens rostral division anterior part; REd, reuniens rostral division dorsal part; REl, reuniens rostral division lateral part; REm, reuniens rostral division median part; REv, reuniens rostral division ventral part; REcm, reuniens caudal division median part; REcd, reuniens caudal division dorsal part; REcp, reuniens caudal division posterior part.

    Journal: bioRxiv

    Article Title: Calretinin and calbindin architecture of the midline thalamus associated with prefrontal-hippocampal circuitry

    doi: 10.1101/2020.07.21.214973

    Figure Lengend Snippet: Distribution of DAB CR + and DAB CB + cells is not the same across all RE internal subdivisions A: Brightfield images showing the distribution of DAB CR + cells in RE across the rostro-caudal axis of the thalamus. CR + cell area density varied depending on the subdivision of RE in which they were located. B: Distribution of DAB CB + cells. When compared, CR + and CB + cell distribution in all RE’s subregions and across the rostral to caudal levels did not appear to be the same. Overlay shown adapted from Swanson (2018) to highlight all RE internal subdivisions. Scale bar = 100μm. C: Comparison of DAB CR + and DAB CB + cell area density (cells/0.01mm 2 ) in all subdivisions of RE across the rostro-caudal axis. CB + cell densities were higher than CR + cell densities (except REm). Additionally, REl, REv and REcd subdivisions exhibited large CB + cell densities compared to other RE subregions. A moderate size effect was found for CB + cell area density across all levels and RE subdivisions (Hedges’ d=0.32). Abbreviations: β, bregma; CB, calbindin; CR, calretinin; DAB, 3,3’-Diaminobenzidine; PRe, perireuniens, RE, nucleus reuniens of the thalamus, REa, reuniens rostral division anterior part; REd, reuniens rostral division dorsal part; REl, reuniens rostral division lateral part; REm, reuniens rostral division median part; REv, reuniens rostral division ventral part; REcm, reuniens caudal division median part; REcd, reuniens caudal division dorsal part; REcp, reuniens caudal division posterior part.

    Article Snippet: The peroxidase reaction was produced by incubating the sections for 5 to 12 minutes in a DAB substrate solution (SK-4100; Vector Labs, CA).

    Techniques:

    Absence of basolateral hephaestin (Hp) on duodenal enterocytes from sex linked anaemia ( sla ) mice. 3,3′-Diaminobenzidine immunohistochemistry of duodenal sections using an antiserum to the C terminus of Hp. (A) 100× of C57BL/6J (WT) duodenal sections; (B) 3× magnification of boxed area in (A). Arrows indicate lateral (L) and supranuclear (SN) staining. (C) 100× of sla duodenal sections; (D) 3× magnification of boxed area in (C). Arrows indicate supranuclear (SN) staining. There was no appreciable lateral staining.

    Journal: Gut

    Article Title: Mislocalisation of hephaestin, a multicopper ferroxidase involved in basolateral intestinal iron transport, in the sex linked anaemia mouse

    doi: 10.1136/gut.2003.019026

    Figure Lengend Snippet: Absence of basolateral hephaestin (Hp) on duodenal enterocytes from sex linked anaemia ( sla ) mice. 3,3′-Diaminobenzidine immunohistochemistry of duodenal sections using an antiserum to the C terminus of Hp. (A) 100× of C57BL/6J (WT) duodenal sections; (B) 3× magnification of boxed area in (A). Arrows indicate lateral (L) and supranuclear (SN) staining. (C) 100× of sla duodenal sections; (D) 3× magnification of boxed area in (C). Arrows indicate supranuclear (SN) staining. There was no appreciable lateral staining.

    Article Snippet: Staining was visualised with 3,3′-diaminobenzidine (DAB substrate kit; Vector Laboratories) and counterstained with Gill’s haematoxylin No 2 (Polysciences Inc., Warrington, Pennsylvania, USA) including the nickel solution for Hp staining.

    Techniques: Mouse Assay, Immunohistochemistry, Staining

    Basolateral Ireg1 in duodenal enterocytes from sla mice. 3,3′-Diaminobenzidine immunohistochemistry of duodenal sections using an antiserum to Ireg1. (A) 100× of C57BL/6J (WT) duodenal sections; (B) 3× magnification of boxed area in (A). Arrows indicate lateral (L) staining. (C) 100× of sla duodenal sections; (D) 3× magnification of boxed area in (C).

    Journal: Gut

    Article Title: Mislocalisation of hephaestin, a multicopper ferroxidase involved in basolateral intestinal iron transport, in the sex linked anaemia mouse

    doi: 10.1136/gut.2003.019026

    Figure Lengend Snippet: Basolateral Ireg1 in duodenal enterocytes from sla mice. 3,3′-Diaminobenzidine immunohistochemistry of duodenal sections using an antiserum to Ireg1. (A) 100× of C57BL/6J (WT) duodenal sections; (B) 3× magnification of boxed area in (A). Arrows indicate lateral (L) staining. (C) 100× of sla duodenal sections; (D) 3× magnification of boxed area in (C).

    Article Snippet: Staining was visualised with 3,3′-diaminobenzidine (DAB substrate kit; Vector Laboratories) and counterstained with Gill’s haematoxylin No 2 (Polysciences Inc., Warrington, Pennsylvania, USA) including the nickel solution for Hp staining.

    Techniques: Mouse Assay, Immunohistochemistry, Staining

    Hypoxia is reduced in a 3D spheroid model and is predicted to decline in vivo . (A) EMT6 and FaDu spheroids were treated for 24 h with BEZ235 or BKM120, prior to fixation and sectioning. Sections were DAB-stained for pAKT (Ser473) expression. Representative images are shown for BEZ235 (50 nM) and BKM120 (5 μM). (B) The percentage of DAB-positive pixels in the viable region of the spheroid was reduced with all treatments. (C) Spheroids were treated with 50 nM BEZ235 or 5 μM BKM120 for 24 h, 48 h or 24 h followed by 24 h incubation in drug-free medium (24 h + wash). Hypoxia was assessed by staining central spheroid sections for EF5 (red) with Hoechst as a counterstain (blue). Data shown are representative of two independent experiments, n = 10 spheroids. (D) Mean EF5 fluorescence in spheroids treated for 24 h was reduced in both cell lines tested. (E) To assess whether hypoxia was recoverable, mean EF5 fluorescence in spheroids treated for 24 h was compared with that in spheroids treated for either 48 h or 24 h followed by the 24 h wash in drug-free medium. In cells treated with BEZ235, the 24 h wash period led to a return of the hypoxia signal. (F and G) Simulated distribution of oxygen partial pressure (mmHg) in a representative 1497 μm × 1482 μm vascular network derived from ex vivo samples where oxygen consumption was either 100% (F) or 55% (G) of base level. Highly oxygenated regions are in red, scaling down to poorly oxygenated regions in blue. (H) Average hypoxic fraction (defined as

    Journal: Radiotherapy and Oncology

    Article Title: Regulation of O2 consumption by the PI3K and mTOR pathways contributes to tumor hypoxia

    doi: 10.1016/j.radonc.2014.02.007

    Figure Lengend Snippet: Hypoxia is reduced in a 3D spheroid model and is predicted to decline in vivo . (A) EMT6 and FaDu spheroids were treated for 24 h with BEZ235 or BKM120, prior to fixation and sectioning. Sections were DAB-stained for pAKT (Ser473) expression. Representative images are shown for BEZ235 (50 nM) and BKM120 (5 μM). (B) The percentage of DAB-positive pixels in the viable region of the spheroid was reduced with all treatments. (C) Spheroids were treated with 50 nM BEZ235 or 5 μM BKM120 for 24 h, 48 h or 24 h followed by 24 h incubation in drug-free medium (24 h + wash). Hypoxia was assessed by staining central spheroid sections for EF5 (red) with Hoechst as a counterstain (blue). Data shown are representative of two independent experiments, n = 10 spheroids. (D) Mean EF5 fluorescence in spheroids treated for 24 h was reduced in both cell lines tested. (E) To assess whether hypoxia was recoverable, mean EF5 fluorescence in spheroids treated for 24 h was compared with that in spheroids treated for either 48 h or 24 h followed by the 24 h wash in drug-free medium. In cells treated with BEZ235, the 24 h wash period led to a return of the hypoxia signal. (F and G) Simulated distribution of oxygen partial pressure (mmHg) in a representative 1497 μm × 1482 μm vascular network derived from ex vivo samples where oxygen consumption was either 100% (F) or 55% (G) of base level. Highly oxygenated regions are in red, scaling down to poorly oxygenated regions in blue. (H) Average hypoxic fraction (defined as

    Article Snippet: To assess signaling inhibition in spheroids, sections were stained with anti-pAKT antibody using ImmPRESS™ reagent kit (MP-7401, VectorLabs) and DAB Peroxidase substrate kit (SK-4100, VectorLabs).

    Techniques: In Vivo, Staining, Expressing, Incubation, Fluorescence, Derivative Assay, Ex Vivo

    Dysregulated IRF expression in patients with ductal carcinoma . A . Normal and ADH breast tissue specimens were stained by IF or IHC. Antibodies recognizing IRF5 (FITC), IRF1 (Cy3) and DAPI for the nucleus were used for IF. For IHC, tissues were stained for IRF1 with DAB (brownish-red), IRF5 with BAP (blue), and nucleus with Fast Red mounting buffer. B . Same as in (A), except tissue samples from patients with ADH were stained by IF with IRF5 (FITC) and CK14 (Cy3) in order to confirm expression of IRF5 in myoepithelial cells. C . Same as in (A), except tissues from patients with DCIS and IDC were examined. Representative pictures of low grade and high grade DCIS are shown illustrating distinct differences between IRF1 and IRF5 expression. Images were taken on a Zeiss Axiovert Apotome microscope at 20 × or 40 × magnification. Scale bars are 50 μm.

    Journal: Breast Cancer Research : BCR

    Article Title: Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis

    doi: 10.1186/bcr3053

    Figure Lengend Snippet: Dysregulated IRF expression in patients with ductal carcinoma . A . Normal and ADH breast tissue specimens were stained by IF or IHC. Antibodies recognizing IRF5 (FITC), IRF1 (Cy3) and DAPI for the nucleus were used for IF. For IHC, tissues were stained for IRF1 with DAB (brownish-red), IRF5 with BAP (blue), and nucleus with Fast Red mounting buffer. B . Same as in (A), except tissue samples from patients with ADH were stained by IF with IRF5 (FITC) and CK14 (Cy3) in order to confirm expression of IRF5 in myoepithelial cells. C . Same as in (A), except tissues from patients with DCIS and IDC were examined. Representative pictures of low grade and high grade DCIS are shown illustrating distinct differences between IRF1 and IRF5 expression. Images were taken on a Zeiss Axiovert Apotome microscope at 20 × or 40 × magnification. Scale bars are 50 μm.

    Article Snippet: The second staining was with 1:200 diluted anti-IRF1, Peroxidase anti-Rabbit IgG (Vector Laboratories, PI-1000) and developed with DAB Substrate Kit (Vector Laboratories, SK-4100).

    Techniques: Expressing, Staining, Immunohistochemistry, Microscopy

    Immunohistochemical staining of claudin-7 expression in (A and B) normal endometrium and (C and D) cancer tissues. Original magnification, (A and C) ×200 and (B and D) ×400; staining, DAB. (E) Claudin-7 mRNA expression in endometrial cancer cell lines, as detected by real-time reverse trancription-polymerase chain reaction analysis. N, normal tissues; T, tumor tissues; DAB, 3,3′-diaminobenzidine.

    Journal: Oncology Letters

    Article Title: Downregulation of claudin-7 potentiates cellular proliferation and invasion in endometrial cancer

    doi: 10.3892/ol.2013.1330

    Figure Lengend Snippet: Immunohistochemical staining of claudin-7 expression in (A and B) normal endometrium and (C and D) cancer tissues. Original magnification, (A and C) ×200 and (B and D) ×400; staining, DAB. (E) Claudin-7 mRNA expression in endometrial cancer cell lines, as detected by real-time reverse trancription-polymerase chain reaction analysis. N, normal tissues; T, tumor tissues; DAB, 3,3′-diaminobenzidine.

    Article Snippet: Antibody staining was visualized with 3,3′-diaminobenzidine (DAB; Invitrogen Life Technologies, Carlsbad, CA, USA).

    Techniques: Immunohistochemistry, Staining, Expressing, Polymerase Chain Reaction

    Cellular Organization of the Adult Thoracic NG (A–C) Adult VNCs immunostained with anti-BRP (neuropil marker, blue) (A1–C) and anti-Dll (NG marker, green) (B) with astrocytes expressing mCD8::GFP (membrane marker, green) (A1 and A2), H2B::RFP (nuclear marker, red) (B) or the multicolor system FB1.1 (single-cell marker, red, yellow, and green) (C) under the control of alrm-Gal4 . (B) Astrocyte (H2B::RFP + , Dll + ): arrow, EG (Dll + ): arrowhead. . ) (F) under the control of R56F03-Gal4 . (E) Astrocyte: arrow (Dll + ), EG (H2B::RFP + , Dll + ): arrowhead. (D) arrow: in more than half of the samples analyzed (n = 10) a cell body of an EG (GFP + , Dll + ) was observed inside the neuropil, next to axon bundles. . (G and H) Prothoracic neuromeres with EG-expressing mCD4::GFP and immunostained with anti-BRP (blue) and anti-Elav (neuron marker, red) (G) or anti-NCad (neuropil marker, red) and Nrg (axon marker, blue) (H). (G) Asterisk: Elav + . (I and J) Prothoracic neuromeres with EG-expressing mCD4::GFP (I) and GFP::mCD8::HRP (J) immunostained with anti-BRP (blue) (I) or labeled with DAB (brown) (J). (K) Low-magnification electron microscope image of the boxed region in (J). (L–O) Enlargement of the boxed regions in (K). (L2) Enlargement of the boxed region in (L1). Note: (L1) and (L2) show the organization of the EG processes around the neuropil while (M)–(O) show the EG processes inside the neuropil. Pink lines outline axons. PG, perineurial glia; SPG, subperineurial glia; NG, neuropil glia; C, cortex; Np, neuropil; NL, neural lamella. (P) Bottom, schematic of adult CNS (blue: neuropils, gray: cortex); top, single astrocyte and EG labeled with mCD8::GFP under the control of alrm-Gal4 and R56F03-Gal4 using MARCM. Axes: green (posterior), blue (dorsal), red (medial). (Q) Average number of NG in the adult VNC, in which EG or astrocytes were expressing H2B::RFP under the control of R56F03-Gal4 or alrm-Gal4 , respectively, and immunostained with anti-Dll. Number of samples = 4/genotype. Error bars indicate SD. ProNm, Prothoracic neuromere; AMesoNm, Accessory mesothoracic neuromere; MesoNm, Mesothoracic neuromere; MetaNm, Metathoracic neuromere; ANm, Abdominal neuromeres; FS, frontal cross-section; TS, transverse cross-section; 3D, 3-dimensional reconstruction of confocal image stack; pMP, partial maximum projection.

    Journal: Neuron

    Article Title: Differing Strategies Despite Shared Lineages of Motor Neurons and Glia to Achieve Robust Development of an Adult Neuropil in Drosophila

    doi: 10.1016/j.neuron.2018.01.007

    Figure Lengend Snippet: Cellular Organization of the Adult Thoracic NG (A–C) Adult VNCs immunostained with anti-BRP (neuropil marker, blue) (A1–C) and anti-Dll (NG marker, green) (B) with astrocytes expressing mCD8::GFP (membrane marker, green) (A1 and A2), H2B::RFP (nuclear marker, red) (B) or the multicolor system FB1.1 (single-cell marker, red, yellow, and green) (C) under the control of alrm-Gal4 . (B) Astrocyte (H2B::RFP + , Dll + ): arrow, EG (Dll + ): arrowhead. . ) (F) under the control of R56F03-Gal4 . (E) Astrocyte: arrow (Dll + ), EG (H2B::RFP + , Dll + ): arrowhead. (D) arrow: in more than half of the samples analyzed (n = 10) a cell body of an EG (GFP + , Dll + ) was observed inside the neuropil, next to axon bundles. . (G and H) Prothoracic neuromeres with EG-expressing mCD4::GFP and immunostained with anti-BRP (blue) and anti-Elav (neuron marker, red) (G) or anti-NCad (neuropil marker, red) and Nrg (axon marker, blue) (H). (G) Asterisk: Elav + . (I and J) Prothoracic neuromeres with EG-expressing mCD4::GFP (I) and GFP::mCD8::HRP (J) immunostained with anti-BRP (blue) (I) or labeled with DAB (brown) (J). (K) Low-magnification electron microscope image of the boxed region in (J). (L–O) Enlargement of the boxed regions in (K). (L2) Enlargement of the boxed region in (L1). Note: (L1) and (L2) show the organization of the EG processes around the neuropil while (M)–(O) show the EG processes inside the neuropil. Pink lines outline axons. PG, perineurial glia; SPG, subperineurial glia; NG, neuropil glia; C, cortex; Np, neuropil; NL, neural lamella. (P) Bottom, schematic of adult CNS (blue: neuropils, gray: cortex); top, single astrocyte and EG labeled with mCD8::GFP under the control of alrm-Gal4 and R56F03-Gal4 using MARCM. Axes: green (posterior), blue (dorsal), red (medial). (Q) Average number of NG in the adult VNC, in which EG or astrocytes were expressing H2B::RFP under the control of R56F03-Gal4 or alrm-Gal4 , respectively, and immunostained with anti-Dll. Number of samples = 4/genotype. Error bars indicate SD. ProNm, Prothoracic neuromere; AMesoNm, Accessory mesothoracic neuromere; MesoNm, Mesothoracic neuromere; MetaNm, Metathoracic neuromere; ANm, Abdominal neuromeres; FS, frontal cross-section; TS, transverse cross-section; 3D, 3-dimensional reconstruction of confocal image stack; pMP, partial maximum projection.

    Article Snippet: HRP expression was revealed overnight using the DAB substrate kit (Thermofisher).

    Techniques: Marker, Expressing, Labeling, Microscopy

    Avulsion of dorsal roots dramatically disrupted the dorsal spinal cord, causing an influx of blood cells only at the lesion site, while rhizotomy did not result in blood cell influx. Incubation of tissue with 3′,3-diaminobenzidine (DAB) revealed

    Journal: Journal of Neurotrauma

    Article Title: Below Level Central Pain Induced by Discrete Dorsal Spinal Cord Injury

    doi: 10.1089/neu.2010.1311

    Figure Lengend Snippet: Avulsion of dorsal roots dramatically disrupted the dorsal spinal cord, causing an influx of blood cells only at the lesion site, while rhizotomy did not result in blood cell influx. Incubation of tissue with 3′,3-diaminobenzidine (DAB) revealed

    Article Snippet: For one, we took advantage of the interaction between 3′,3-diaminobenzidine (DAB; Sigma-Aldrich, St. Louis, MO), and endogenous peroxidases expressed by red blood cells.

    Techniques: Incubation

    Carboxymethyl cellulose-coated carbon nanotubes (CMC-CNTs) stably bind to purified human properdin and recombinant TSR4+5, as shown via SDS-PAGE (A) and western blot (B) . (A) CMC-CNTs were incubated with recombinant human properdin, or thrombospondin type I repeat (TSR) 4 + 5-MBP fusion protein and BSA overnight in the affinity buffer. Carbon nanotubes (CNTs) were washed extensively via centrifugation and run on an SDS-PAGE (12%) under reduced conditions. Properdin migrated at ~55 kDa and MBP-TSR4+5 migrated as a single band at ~53 kDa. (B) In parallel, the SDS-PAGE was transferred onto nitrocellulose membrane for 2 h at 320 mA. The blot was probed with anti-human properdin (polyclonal) antibodies, followed by protein A–HRP conjugate and developed using 3,3′-diaminobenzidine.

    Journal: Frontiers in Immunology

    Article Title: Human Properdin Opsonizes Nanoparticles and Triggers a Potent Pro-inflammatory Response by Macrophages without Involving Complement Activation

    doi: 10.3389/fimmu.2018.00131

    Figure Lengend Snippet: Carboxymethyl cellulose-coated carbon nanotubes (CMC-CNTs) stably bind to purified human properdin and recombinant TSR4+5, as shown via SDS-PAGE (A) and western blot (B) . (A) CMC-CNTs were incubated with recombinant human properdin, or thrombospondin type I repeat (TSR) 4 + 5-MBP fusion protein and BSA overnight in the affinity buffer. Carbon nanotubes (CNTs) were washed extensively via centrifugation and run on an SDS-PAGE (12%) under reduced conditions. Properdin migrated at ~55 kDa and MBP-TSR4+5 migrated as a single band at ~53 kDa. (B) In parallel, the SDS-PAGE was transferred onto nitrocellulose membrane for 2 h at 320 mA. The blot was probed with anti-human properdin (polyclonal) antibodies, followed by protein A–HRP conjugate and developed using 3,3′-diaminobenzidine.

    Article Snippet: Protein A–horseradish peroxidase (1:1,000; Thermo Scientific) in PBS was added and left at room temperature for 1 h. The blot was washed again with PBST three times, and the color was developed using 3,3′-diaminobenzidine (DAB) (Sigma-Aldrich).

    Techniques: Stable Transfection, Purification, Recombinant, SDS Page, Western Blot, Incubation, Centrifugation

    In situ detection of H 2 O 2 using 3,3′-diaminobenzidine (DAB) staining on rubber tree leaves after treatment with 5 mM SA. The rubber tree leaves were sprayed with either DW as a control or 5 mM SA. Leaf pieces were collected and stained after treatment at different time points (0, 3, 6, 12, 24, 48, 72, 96 and 120 h). Pictures represent three independent biological replicates. The dark brown precipitates indicate the presence and distribution of H 2 O 2 in plant cells, which could be visualized using a light microscope (scale bars = 100 μm).

    Journal: International Journal of Molecular Sciences

    Article Title: Salicylic Acid Induces Resistance in Rubber Tree against Phytophthora palmivora

    doi: 10.3390/ijms19071883

    Figure Lengend Snippet: In situ detection of H 2 O 2 using 3,3′-diaminobenzidine (DAB) staining on rubber tree leaves after treatment with 5 mM SA. The rubber tree leaves were sprayed with either DW as a control or 5 mM SA. Leaf pieces were collected and stained after treatment at different time points (0, 3, 6, 12, 24, 48, 72, 96 and 120 h). Pictures represent three independent biological replicates. The dark brown precipitates indicate the presence and distribution of H 2 O 2 in plant cells, which could be visualized using a light microscope (scale bars = 100 μm).

    Article Snippet: In situ detection of H2 O2 content in rubber tree leaf cells was performed by staining with 3, 3′-diaminobenzidine (DAB; Sigma Chem.

    Techniques: In Situ, Staining, Light Microscopy

    MAD1 expression negatively correlates with that of miR-125b in HNOC. ( a ) Reciprocal relation of miR-125b and MAD1 in HNOC tumours. Total RNA was isolated from primary HNOC tissues ( n =25) and adjacent normal tissues ( n =20). cDNA was prepared by stem-loop primers specific for miR-125b and miR-17-5p and subjected to RT-PCR. Values were normalized to those of miR-17-5p. For MAD1, the above RNA was reverse transcribed and cDNA was subjected to RT-PCR using MAD1 -specific primers. GAPDH was used as endogenous control. ΔΔCt values were calculated and data plotted in terms of log 2 of relative expressions. P -values are indicated. ‘ n ' represents the number of tumour samples and circles represent outliers. ( b ) Representative images showing inverse relation between Mad1 and miR-125b in primary HNOC tissues. Paraffin-embedded tissues samples (in which miR-125b expression was found to be low and Mad1 expression was high and vice versa ; n =16) were processed for IHC with antibody against Mad1, stained with 3,3′diaminobenzidine and counterstained with hematoxylin. Nuclei, which stained mild or deep brown represent moderate or high expression of Mad1, respectively. Blue/bluish-purple staining of nuclei represents low Mad1 expression. Images represent × 20 magnification, while inset images represent × 40 magnification. Scale bars represent 50 μ m. T1, T2, T3, T4, T5 and T6 are representative tumour samples taken from six individual patients

    Journal: Cell Death and Differentiation

    Article Title: miR-125b promotes cell death by targeting spindle assembly checkpoint gene MAD1 and modulating mitotic progression

    doi: 10.1038/cdd.2012.135

    Figure Lengend Snippet: MAD1 expression negatively correlates with that of miR-125b in HNOC. ( a ) Reciprocal relation of miR-125b and MAD1 in HNOC tumours. Total RNA was isolated from primary HNOC tissues ( n =25) and adjacent normal tissues ( n =20). cDNA was prepared by stem-loop primers specific for miR-125b and miR-17-5p and subjected to RT-PCR. Values were normalized to those of miR-17-5p. For MAD1, the above RNA was reverse transcribed and cDNA was subjected to RT-PCR using MAD1 -specific primers. GAPDH was used as endogenous control. ΔΔCt values were calculated and data plotted in terms of log 2 of relative expressions. P -values are indicated. ‘ n ' represents the number of tumour samples and circles represent outliers. ( b ) Representative images showing inverse relation between Mad1 and miR-125b in primary HNOC tissues. Paraffin-embedded tissues samples (in which miR-125b expression was found to be low and Mad1 expression was high and vice versa ; n =16) were processed for IHC with antibody against Mad1, stained with 3,3′diaminobenzidine and counterstained with hematoxylin. Nuclei, which stained mild or deep brown represent moderate or high expression of Mad1, respectively. Blue/bluish-purple staining of nuclei represents low Mad1 expression. Images represent × 20 magnification, while inset images represent × 40 magnification. Scale bars represent 50 μ m. T1, T2, T3, T4, T5 and T6 are representative tumour samples taken from six individual patients

    Article Snippet: Slides were developed using 3,3′diaminobenzidine as the chromogen after treating with HRP-conjugated secondary antibody (Sigma) and then counterstained with hematoxylin.

    Techniques: Expressing, Isolation, Reverse Transcription Polymerase Chain Reaction, Immunohistochemistry, Staining

    Immunoaffinity enrichment of Dermanyssus gallinae proteins using immobilised antigen-specific IgY. Yolk-IgY generated against ion exchange chromatography group 4 proteins was cross-linked to a HiTrap N-hydroxysuccinimide (NHS)-activated column and IEX group 4 proteins were selectively bound to the column, washed to remove unbound material and then eluted into an affinity enriched fraction with 0.1 M Glycine, 6 M Urea, pH 2.5. (A) The IEX group 4 proteins prior to affinity purification (lanes 1 and 3) and the eluted affinity-enriched material (lanes 2 and 4) were separated on a 12% Bis–Tris Novex gel (GE Healthcare) and transferred to nitrocellulose. Lanes 1 and 2 of the immunoblot were probed with a 1:200 dilution of sera from naive hens that had not been immunised with the soluble mite extract and lanes 3 and 4 were probed with a 1:200 dilution of post-vaccination sera from hens immunised with IEX group 4. Bound IgY was detected with rabbit anti-IgY-peroxidase (Sigma) and visualised using SIGMA FAST ™ 3,3′-diaminobenzidine substrate (Sigma). (B) The eluted immunoaffinity enriched IEX group 4 material was concentrated fivefold by lyophilisation and 30 μl was electrophoretically separated as before and stained with SimplyBlue™ SafeStain (lane 5). The gel lane was sectioned into 24 equally-sized horizontal slices and the proteins extracted and subjected to liquid chromatography–electrospray ionisation–tandem mass spectrometry (LC–ESI–MS/MS).

    Journal: International Journal for Parasitology

    Article Title: Identification and evaluation of vaccine candidate antigens from the poultry red mite (Dermanyssus gallinae)

    doi: 10.1016/j.ijpara.2015.07.004

    Figure Lengend Snippet: Immunoaffinity enrichment of Dermanyssus gallinae proteins using immobilised antigen-specific IgY. Yolk-IgY generated against ion exchange chromatography group 4 proteins was cross-linked to a HiTrap N-hydroxysuccinimide (NHS)-activated column and IEX group 4 proteins were selectively bound to the column, washed to remove unbound material and then eluted into an affinity enriched fraction with 0.1 M Glycine, 6 M Urea, pH 2.5. (A) The IEX group 4 proteins prior to affinity purification (lanes 1 and 3) and the eluted affinity-enriched material (lanes 2 and 4) were separated on a 12% Bis–Tris Novex gel (GE Healthcare) and transferred to nitrocellulose. Lanes 1 and 2 of the immunoblot were probed with a 1:200 dilution of sera from naive hens that had not been immunised with the soluble mite extract and lanes 3 and 4 were probed with a 1:200 dilution of post-vaccination sera from hens immunised with IEX group 4. Bound IgY was detected with rabbit anti-IgY-peroxidase (Sigma) and visualised using SIGMA FAST ™ 3,3′-diaminobenzidine substrate (Sigma). (B) The eluted immunoaffinity enriched IEX group 4 material was concentrated fivefold by lyophilisation and 30 μl was electrophoretically separated as before and stained with SimplyBlue™ SafeStain (lane 5). The gel lane was sectioned into 24 equally-sized horizontal slices and the proteins extracted and subjected to liquid chromatography–electrospray ionisation–tandem mass spectrometry (LC–ESI–MS/MS).

    Article Snippet: Bound IgY was detected with anti-IgY-peroxidase conjugate (Sigma) and visualised using SIGMA FAST ™ 3,3′-diaminobenzidine substrate (Sigma).

    Techniques: Generated, Ion Exchange Chromatography, Affinity Purification, Staining, Liquid Chromatography, Mass Spectrometry

    Immunoreactive profiles of soluble mite extract of Dermanyssus gallinae separated by ion exchange chromatography. Ion exchange chromatography fractions were pooled into five groups (IEX groups 1–5), with IEX group 1 consisting of the column flow-through (FT) and IEX groups 2–5 of selectively pooled eluted fractions. Two microgram of IEX groups 1–5 (lanes 1–5) and 2 μg of the soluble mite extract (SME, lane 6) were separated on a 12% Bis–Tris Novex gel (GE Healthcare), transferred to nitrocellulose and probed with IgY generated against the soluble mite extract (A) or IgY from a hen that had not been injected with soluble mite extract (B). Bound IgY was detected with rabbit anti-IgY-peroxidase (Sigma) and visualised with SIGMA FAST ™ 3,3′-diaminobenzidine substrate (Sigma).

    Journal: International Journal for Parasitology

    Article Title: Identification and evaluation of vaccine candidate antigens from the poultry red mite (Dermanyssus gallinae)

    doi: 10.1016/j.ijpara.2015.07.004

    Figure Lengend Snippet: Immunoreactive profiles of soluble mite extract of Dermanyssus gallinae separated by ion exchange chromatography. Ion exchange chromatography fractions were pooled into five groups (IEX groups 1–5), with IEX group 1 consisting of the column flow-through (FT) and IEX groups 2–5 of selectively pooled eluted fractions. Two microgram of IEX groups 1–5 (lanes 1–5) and 2 μg of the soluble mite extract (SME, lane 6) were separated on a 12% Bis–Tris Novex gel (GE Healthcare), transferred to nitrocellulose and probed with IgY generated against the soluble mite extract (A) or IgY from a hen that had not been injected with soluble mite extract (B). Bound IgY was detected with rabbit anti-IgY-peroxidase (Sigma) and visualised with SIGMA FAST ™ 3,3′-diaminobenzidine substrate (Sigma).

    Article Snippet: Bound IgY was detected with anti-IgY-peroxidase conjugate (Sigma) and visualised using SIGMA FAST ™ 3,3′-diaminobenzidine substrate (Sigma).

    Techniques: Ion Exchange Chromatography, Flow Cytometry, Generated, Injection

    Representative images of GFAP+ astrocytes in the hippocampus of adult Long-Evans hooded male rats exposed to low-F − chow/RO-H 2 O or low-F − chow/20 ppm F − drinking water beginning on gestational day 6. ( a ) Suprapyramidal blade of the dentate gyrus. ( b ) CA1 pyramidal cell layer. Cells displayed normal process-bearing morphology with no evidence of hypertrophy. 3,3-diaminobenzidine staining (brown). Hematoxylin counterstain (blue) showed no disruption of the normal morphology of the hippocampal regions and no evidence of neuronal death. ( n = 6). Scale bar = 100 μm

    Journal: Neurotoxicity Research

    Article Title: An Evaluation of Neurotoxicity Following Fluoride Exposure from Gestational Through Adult Ages in Long-Evans Hooded Rats

    doi: 10.1007/s12640-018-9870-x

    Figure Lengend Snippet: Representative images of GFAP+ astrocytes in the hippocampus of adult Long-Evans hooded male rats exposed to low-F − chow/RO-H 2 O or low-F − chow/20 ppm F − drinking water beginning on gestational day 6. ( a ) Suprapyramidal blade of the dentate gyrus. ( b ) CA1 pyramidal cell layer. Cells displayed normal process-bearing morphology with no evidence of hypertrophy. 3,3-diaminobenzidine staining (brown). Hematoxylin counterstain (blue) showed no disruption of the normal morphology of the hippocampal regions and no evidence of neuronal death. ( n = 6). Scale bar = 100 μm

    Article Snippet: Sections were incubated with rabbit anti-cow glial fibrillary acidic protein (Dako GFAP; 1:7000; RT; 30 min; Agilent Technologies, Carpinteria, CA) then incubated with biotinylated goat anti-rabbit IgG (1:500; Vector-Labs) and detected with Vectastain Elite ABC R.T.U. (Vector Labs); 3,3-diaminobenzidine (DAB, Agilent Technologies).

    Techniques: Staining

    Representative images of Iba-1+ microglia in the hippocampus of adult Long-Evans hooded rats exposed to low-F − chow/RO-H 2 O or low-F − chow/20 ppm F − drinking water beginning on gestational day 6. ( a ) Suprapyramidal blade of the dentate gyrus. ( b ) CA1 pyramidal cell layer. Cells displayed normal process-bearing morphology with no evidence of reactivity or activation. 3,3-diaminobenzidine staining (brown). Hematoxylin counterstain (blue) ( n = 6). Scale bar = 100 μm

    Journal: Neurotoxicity Research

    Article Title: An Evaluation of Neurotoxicity Following Fluoride Exposure from Gestational Through Adult Ages in Long-Evans Hooded Rats

    doi: 10.1007/s12640-018-9870-x

    Figure Lengend Snippet: Representative images of Iba-1+ microglia in the hippocampus of adult Long-Evans hooded rats exposed to low-F − chow/RO-H 2 O or low-F − chow/20 ppm F − drinking water beginning on gestational day 6. ( a ) Suprapyramidal blade of the dentate gyrus. ( b ) CA1 pyramidal cell layer. Cells displayed normal process-bearing morphology with no evidence of reactivity or activation. 3,3-diaminobenzidine staining (brown). Hematoxylin counterstain (blue) ( n = 6). Scale bar = 100 μm

    Article Snippet: Sections were incubated with rabbit anti-cow glial fibrillary acidic protein (Dako GFAP; 1:7000; RT; 30 min; Agilent Technologies, Carpinteria, CA) then incubated with biotinylated goat anti-rabbit IgG (1:500; Vector-Labs) and detected with Vectastain Elite ABC R.T.U. (Vector Labs); 3,3-diaminobenzidine (DAB, Agilent Technologies).

    Techniques: Activation Assay, Staining

    Both β-catenin and Ras levels are increased in adenocarcinoma, metastatic adenocarcinoma and tumor budding in colon cancer TMA specimens of normal mucosa, adenocarcinoma, metastatic adenocarcinoma, or tumor buddings were subjected to the IHC analyses by using β-catenin or Ras antibody followed by 3, 3'-diaminobenzidine (DAB) staining. A. Representative image of IHC analyses for β-catenin and Ras in normal tissues and at different stages of colorectal tumorigenesis. Boxes indicate the enlarged areas. Scale bar= 50μm. B., D. Quantitative analyses of positive signal were performed by comparing the H-Scores of staining for β-catenin and Ras in TMA samples. Normal Mucosa (n=2), Adenocarcinoma (n=26), Metastatic adenocarcinoma (n=24), Tumor budding (n=7). C. Representative images of IHC analyses for β-catenin and Ras in tumor budding compared with a neighboring adenocarcinoma in colon cancer TMA samples. The yellow boxes represent enlarged in the right panel. Scale bar= 50μm. D. Quantitative analysis for β-catenin and Ras was performed by comparing tumor budding with neighboring adenocarcinoma based on H-Score (n=10). Two-sided Student t test was used to determine statistical significance using GraphPad Prism5 Software. Error bars represent 95% confidence intervals.

    Journal: Oncotarget

    Article Title: KY1022, a small molecule destabilizing Ras via targeting the Wnt/β-catenin pathway, inhibits development of metastatic colorectal cancer

    doi: 10.18632/oncotarget.13172

    Figure Lengend Snippet: Both β-catenin and Ras levels are increased in adenocarcinoma, metastatic adenocarcinoma and tumor budding in colon cancer TMA specimens of normal mucosa, adenocarcinoma, metastatic adenocarcinoma, or tumor buddings were subjected to the IHC analyses by using β-catenin or Ras antibody followed by 3, 3'-diaminobenzidine (DAB) staining. A. Representative image of IHC analyses for β-catenin and Ras in normal tissues and at different stages of colorectal tumorigenesis. Boxes indicate the enlarged areas. Scale bar= 50μm. B., D. Quantitative analyses of positive signal were performed by comparing the H-Scores of staining for β-catenin and Ras in TMA samples. Normal Mucosa (n=2), Adenocarcinoma (n=26), Metastatic adenocarcinoma (n=24), Tumor budding (n=7). C. Representative images of IHC analyses for β-catenin and Ras in tumor budding compared with a neighboring adenocarcinoma in colon cancer TMA samples. The yellow boxes represent enlarged in the right panel. Scale bar= 50μm. D. Quantitative analysis for β-catenin and Ras was performed by comparing tumor budding with neighboring adenocarcinoma based on H-Score (n=10). Two-sided Student t test was used to determine statistical significance using GraphPad Prism5 Software. Error bars represent 95% confidence intervals.

    Article Snippet: The samples were then incubated in ABC kit (Vector Laboratories) for 1 hour, stained with 3, 3′-diaminobenzidine (DAB; Dako) for 3−7 minutes and counterstained with Mayer's hematoxylin (Muto).

    Techniques: Immunohistochemistry, Staining, Software

    Detection of ROS and expression of ROS marker genes in inbreds with contrasting levels of submergence tolerance. (a.) Detection of H 2 O 2 and superoxide in selected submergence tolerant and sensitive maize genotypes. A subset of four nested association mapping panel parents were selected based on visual leaf scoring for ROS staining assays after submergence. Hydrogen peroxide was detected using stain 3,3′-diaminobenzidine tetrahydrochloride (DAB), while superoxide was detected using nitroblue tetrazolium (NBT). Both assays were performed after 96 h of submergence. In B97, the third leaf was barely emerged and not used for the assay. (b-d) Real-time quantitative PCR of ROS genes. ROS genes were selected based on the study by Gadjev, Vanderauwera and Gechev [ 30 ] and showed a significant induction in response to at least two stresses that induce ROS formation in Arabidopsis. (b.) ALTERNATIVE OXIDASE 1a ( AOX1a ); (c.) WRKY6 (d.) CYTOCHROME P81 D8 ( CYP81D8 ). All sample were collected at 24 h and 72 h after submergence. Expression levels are relative to shoot tissue of control plants at 24 h. Letters above bars indicate nonsignificant differences in expression determined using Tukey’s HSD ( p

    Journal: PLoS ONE

    Article Title: Genetic and Molecular Characterization of Submergence Response Identifies Subtol6 as a Major Submergence Tolerance Locus in Maize

    doi: 10.1371/journal.pone.0120385

    Figure Lengend Snippet: Detection of ROS and expression of ROS marker genes in inbreds with contrasting levels of submergence tolerance. (a.) Detection of H 2 O 2 and superoxide in selected submergence tolerant and sensitive maize genotypes. A subset of four nested association mapping panel parents were selected based on visual leaf scoring for ROS staining assays after submergence. Hydrogen peroxide was detected using stain 3,3′-diaminobenzidine tetrahydrochloride (DAB), while superoxide was detected using nitroblue tetrazolium (NBT). Both assays were performed after 96 h of submergence. In B97, the third leaf was barely emerged and not used for the assay. (b-d) Real-time quantitative PCR of ROS genes. ROS genes were selected based on the study by Gadjev, Vanderauwera and Gechev [ 30 ] and showed a significant induction in response to at least two stresses that induce ROS formation in Arabidopsis. (b.) ALTERNATIVE OXIDASE 1a ( AOX1a ); (c.) WRKY6 (d.) CYTOCHROME P81 D8 ( CYP81D8 ). All sample were collected at 24 h and 72 h after submergence. Expression levels are relative to shoot tissue of control plants at 24 h. Letters above bars indicate nonsignificant differences in expression determined using Tukey’s HSD ( p

    Article Snippet: For H2 O2 visualization, leaf tissue was collected as described above, immersed in 1 mg/mL 3,3-diaminobenzidine tetrahydrochloride (DAB; Sigma-Aldrich, St. Louis, MO, USA) in 50 mM tris-acetate buffer (pH 5.0), and were incubated at 25ºC for 24 h in complete darkness.

    Techniques: Expressing, Marker, Staining, Real-time Polymerase Chain Reaction

    Immunohistochemical analysis by MsMab-1 against tissue microarray of osteosarcomas. Osteosarcoma tissue microarray was stained with MsMab-1 followed by LSAB kit. Color was developed using DAB, and was counterstained with hematoxylin. Typical results were shown: (A) No. 3, (B) No. 5, (C) No. 13, (D) No. 15, (E) No. 18, (F) No. 24, (G) No. 26, (H) No. 31, and (I) No. 32. Insets show that MsMab-1 stained cytoplasm (A–I). Magnification: 200×. MsMab, multi-specific anti-mutated IDH1/2 mAb; DAB, 3,3-diaminobenzidine tetrahydrochloride.

    Journal: Cancer Medicine

    Article Title: Isocitrate dehydrogenase 2 mutation is a frequent event in osteosarcoma detected by a multi-specific monoclonal antibody MsMab-1

    doi: 10.1002/cam4.149

    Figure Lengend Snippet: Immunohistochemical analysis by MsMab-1 against tissue microarray of osteosarcomas. Osteosarcoma tissue microarray was stained with MsMab-1 followed by LSAB kit. Color was developed using DAB, and was counterstained with hematoxylin. Typical results were shown: (A) No. 3, (B) No. 5, (C) No. 13, (D) No. 15, (E) No. 18, (F) No. 24, (G) No. 26, (H) No. 31, and (I) No. 32. Insets show that MsMab-1 stained cytoplasm (A–I). Magnification: 200×. MsMab, multi-specific anti-mutated IDH1/2 mAb; DAB, 3,3-diaminobenzidine tetrahydrochloride.

    Article Snippet: Color was developed using 3,3-diaminobenzidine tetrahydrochloride (DAB; Dako) for 10 min, and counterstained with hematoxylin.

    Techniques: Immunohistochemistry, Microarray, Staining

    Representative images (20x magnification) of airway sections stained for cathepsins. Immunostaining of cathepsins and corresponding isotype controls for cathepsins D (A–D), H (E–H) and K (I–L) from non-diseased and asthmatic sections. Specific staining was detected using a chemical chromophore DAB (brown) and cell nucleus was counterstained with haematoxylin (blue). Abbreviations DAB = 3,3′-diaminobenzidine.

    Journal: PLoS ONE

    Article Title: The Expression and Activity of Cathepsins D, H and K in Asthmatic Airways

    doi: 10.1371/journal.pone.0057245

    Figure Lengend Snippet: Representative images (20x magnification) of airway sections stained for cathepsins. Immunostaining of cathepsins and corresponding isotype controls for cathepsins D (A–D), H (E–H) and K (I–L) from non-diseased and asthmatic sections. Specific staining was detected using a chemical chromophore DAB (brown) and cell nucleus was counterstained with haematoxylin (blue). Abbreviations DAB = 3,3′-diaminobenzidine.

    Article Snippet: Tissue staining was then visualized with substrate chromogen, liquid 3,3′-diaminobenzidine (DAB) (DakoCytomation, Glostrup, CA).

    Techniques: Staining, Immunostaining

    Sections from control mice that underwent direct staining (i.e., no deparaffinization) revealed staining of nuclei (and sometimes distinct cytoplasmic staining) of neurons, myelin, and neuropil staining. (a and b) Staining in the cortex is revealed in nuclei, cytoplasm, and neuropil. (c) The hippocampal CA1 neurons are densely stained. Splotches of staining are apparent in the cortex and hippocampus. (d) Myelin staining in the globus pallidus (right) and caudate or putamen (left) is apparent among staining of cells (dark dots) and neuropil across the field. Some splotches of staining are also apparent (lower left). (e) Dark staining of myelin and neuropil is present in the substantia nigra. Staining of cells (dark dots) across the section is also apparent. (f) Eliminating the potassium ferrocyanide from the staining solution reveals an absence of labeling. Hippocampus is shown (compare to c). Note, some tissue artifact (upper right) is lightly labeled showing that the section was exposed to the DAB solution, and an extremely light depiction of the CA1 neurons can be made out. (g) Staining of the cerebellum reveals staining of granule cells, Purkinje neurons, and staining of neuropil and neurons in the molecular layer, which also has a heterogeneous pattern of staining. Bar in a = 20 µ (a,b). Bar in c = 100 µ (c–g). Note . DAB = 3,3′-diaminobenzidine tetrahydrochloride.

    Journal: ASN NEURO

    Article Title: Enhanced Histochemical Detection of Iron in Paraffin Sections of Mouse Central Nervous System Tissue

    doi: 10.1177/1759091416670978

    Figure Lengend Snippet: Sections from control mice that underwent direct staining (i.e., no deparaffinization) revealed staining of nuclei (and sometimes distinct cytoplasmic staining) of neurons, myelin, and neuropil staining. (a and b) Staining in the cortex is revealed in nuclei, cytoplasm, and neuropil. (c) The hippocampal CA1 neurons are densely stained. Splotches of staining are apparent in the cortex and hippocampus. (d) Myelin staining in the globus pallidus (right) and caudate or putamen (left) is apparent among staining of cells (dark dots) and neuropil across the field. Some splotches of staining are also apparent (lower left). (e) Dark staining of myelin and neuropil is present in the substantia nigra. Staining of cells (dark dots) across the section is also apparent. (f) Eliminating the potassium ferrocyanide from the staining solution reveals an absence of labeling. Hippocampus is shown (compare to c). Note, some tissue artifact (upper right) is lightly labeled showing that the section was exposed to the DAB solution, and an extremely light depiction of the CA1 neurons can be made out. (g) Staining of the cerebellum reveals staining of granule cells, Purkinje neurons, and staining of neuropil and neurons in the molecular layer, which also has a heterogeneous pattern of staining. Bar in a = 20 µ (a,b). Bar in c = 100 µ (c–g). Note . DAB = 3,3′-diaminobenzidine tetrahydrochloride.

    Article Snippet: Sodium azide, sodium borohydride, proteinase K, phosphate-buffered saline (PBS), Trizma base, and 3,3′-diaminobenzidine tetrahydrochloride (DAB) (Cat.#D5905) were purchased from Sigma (St. Louis, MO).

    Techniques: Mouse Assay, Staining, Labeling

    Matched sections from control animals were processed with the (a and b) absence of azide (with hydrogen peroxide) or (c and d) in the presence of azide (no hydrogen peroxide) in methanol. Sections that were processed with deparaffinization, downgraded ethanols, and hydration gave greater staining with azide (c) compared with the absence of azide (a). The labeling at the GP and the SN was greater in sections processed with azide. In sections that were directly stained, azide resulted in greater labeling of cerebellar structures (d) compared with no azide (b). Bar in a = 250 µ (a–d). (e) Solutions containing ferric iron, potassium ferrocyanide, HCl, hydrogen peroxide with (Tubes 64 and 70) and without (Tubes 65 and 71) azide were incubated with DAB for 1 hr (see Methods section). Solutions with azide gave a darker brown solution than solutions without azide. Note . GP = globus pallidus; SN = substantia nigra; DAB = 3,3′-diaminobenzidine tetrahydrochloride.

    Journal: ASN NEURO

    Article Title: Enhanced Histochemical Detection of Iron in Paraffin Sections of Mouse Central Nervous System Tissue

    doi: 10.1177/1759091416670978

    Figure Lengend Snippet: Matched sections from control animals were processed with the (a and b) absence of azide (with hydrogen peroxide) or (c and d) in the presence of azide (no hydrogen peroxide) in methanol. Sections that were processed with deparaffinization, downgraded ethanols, and hydration gave greater staining with azide (c) compared with the absence of azide (a). The labeling at the GP and the SN was greater in sections processed with azide. In sections that were directly stained, azide resulted in greater labeling of cerebellar structures (d) compared with no azide (b). Bar in a = 250 µ (a–d). (e) Solutions containing ferric iron, potassium ferrocyanide, HCl, hydrogen peroxide with (Tubes 64 and 70) and without (Tubes 65 and 71) azide were incubated with DAB for 1 hr (see Methods section). Solutions with azide gave a darker brown solution than solutions without azide. Note . GP = globus pallidus; SN = substantia nigra; DAB = 3,3′-diaminobenzidine tetrahydrochloride.

    Article Snippet: Sodium azide, sodium borohydride, proteinase K, phosphate-buffered saline (PBS), Trizma base, and 3,3′-diaminobenzidine tetrahydrochloride (DAB) (Cat.#D5905) were purchased from Sigma (St. Louis, MO).

    Techniques: Staining, Labeling, Incubation

    Effects of midazolam on ethanol-induced neuroapoptosis in mice. (A) Immunohistochemical analysis of brain sections (n=5/group) was performed using anti-C-3A and was visualized using 3,3′-diaminobenzidine chromogen. The C-3A positive staining was documented under a light microscope and scoring of C-3A-positive cells per mm 2 brain sections was performed. The total number of C-3A-positive cells was presented as the percentage of positive cells compared to the control group. (B) TUNEL staining of brain sections (n=5/group) was performed and analyzed. Data are presented as the mean ± standard deviation. *P

    Journal: Molecular Medicine Reports

    Article Title: Midazolam anesthesia protects neuronal cells from oxidative stress-induced death via activation of the JNK-ERK pathway

    doi: 10.3892/mmr.2016.6031

    Figure Lengend Snippet: Effects of midazolam on ethanol-induced neuroapoptosis in mice. (A) Immunohistochemical analysis of brain sections (n=5/group) was performed using anti-C-3A and was visualized using 3,3′-diaminobenzidine chromogen. The C-3A positive staining was documented under a light microscope and scoring of C-3A-positive cells per mm 2 brain sections was performed. The total number of C-3A-positive cells was presented as the percentage of positive cells compared to the control group. (B) TUNEL staining of brain sections (n=5/group) was performed and analyzed. Data are presented as the mean ± standard deviation. *P

    Article Snippet: Subsequently, tissue sections were washed with TBST and incubated with HRP-labeled anti-rabbit secondary antibody (1:500; cat. no. E0432; Dako Denmark A/S) for 30 min at room temperature followed by visualization with 3,3′-diaminobenzidine chromogen (Dako Denmark A/S).

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Light Microscopy, TUNEL Assay, Standard Deviation

    In situ hybridization of Notch2 and immunohistochemistry of SOX2 in a sagittal section of rat pituitary gland at embryonic day 14.5. a ) Hematoxylin and eosin staining. b ) In situ hybridization of Notch2 . c ) Negative control of in situ hybridization using a sense probe of Notch2 . d ) Double staining of Notch2 detected by in situ hybridization and SOX2 detected by immunohistochemistry. Immunoreactivity for SOX2 was revealed by 3,3'-diaminobenzidine (brown) and the Notch2 in situ hybridization signal with nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate (blue). Arrow: prospective pars intermedia , Double arrow: prospective pars distalis , Triple arrow: prospective pars tuberalis . Bar = 100 μm.

    Journal: Acta Histochemica et Cytochemica

    Article Title: Notch Signaling and Maintenance of SOX2 Expression in Rat Anterior Pituitary Cells

    doi: 10.1267/ahc.17002

    Figure Lengend Snippet: In situ hybridization of Notch2 and immunohistochemistry of SOX2 in a sagittal section of rat pituitary gland at embryonic day 14.5. a ) Hematoxylin and eosin staining. b ) In situ hybridization of Notch2 . c ) Negative control of in situ hybridization using a sense probe of Notch2 . d ) Double staining of Notch2 detected by in situ hybridization and SOX2 detected by immunohistochemistry. Immunoreactivity for SOX2 was revealed by 3,3'-diaminobenzidine (brown) and the Notch2 in situ hybridization signal with nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate (blue). Arrow: prospective pars intermedia , Double arrow: prospective pars distalis , Triple arrow: prospective pars tuberalis . Bar = 100 μm.

    Article Snippet: The ABC method (Vector Laboratories) was performed with 3,3'-diaminobenzidine (Dojindo Laboratories, Kumamoto, Japan) as substrate.

    Techniques: In Situ Hybridization, Immunohistochemistry, Staining, Negative Control, Double Staining

    Immunohistochemistry staining for Notch1- and Notch3-positive cells in lung adenocarcinoma tissues of smokers and non-smokers, as determined using 3,3′-diaminobenzidine and hematoxylin staining (magnification, x200). (A) Notch1-positive cells

    Journal: Oncology Letters

    Article Title: Cigarette smoke induces the expression of Notch3, not Notch1, protein in lung adenocarcinoma

    doi: 10.3892/ol.2015.3329

    Figure Lengend Snippet: Immunohistochemistry staining for Notch1- and Notch3-positive cells in lung adenocarcinoma tissues of smokers and non-smokers, as determined using 3,3′-diaminobenzidine and hematoxylin staining (magnification, x200). (A) Notch1-positive cells

    Article Snippet: 3,3′-diaminobenzidine (DAB; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd.) was used as a chromogen; 100 µl DAB was added to each section and monitored under a microscope until the staining developed, which was followed by immersion slides in dH2 O for 2 × 3 min.

    Techniques: Immunohistochemistry, Staining