dab substrate kit  (Thermo Fisher)


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    DAB Substrate Powder 3 3 diaminobenzidine tetrahydrochloride
    Description:
    Thermo Scientific Pierce DAB enables chromogenic detection of horseradish peroxidase HRP activity based the action of 3 3 diaminobenzidine DAB in Western blot and tissue staining methods DAB 3 3 diaminobenzidine tetrahydrochloride MW 214 1 reacts with HRP in the presence of peroxide to yield an insoluble brown colored product at locations where peroxidase conjugated antibodies are bound to samples The brown precipitate is insoluble in alcohol and other organic solvents making it an excellent substrate for immunohistochemical staining that requires the use of traditional counterstains and mounting media Features of DAB Powder • HRP substrate for detection of horseradish peroxidase activity on solid media including nitrocellulose and PVDF membranes and fixed tissue samples • Chromogenic no special equipment needed for visualization the DAB reaction produces brown bands or spots at sites of reaction with HRP conjugated antibodies or probes • Package options choose powder for custom formulation with buffer and peroxide or a complete kit of preformulated solutions for immediate use DAB has been used in a variety of applications including Western blotting immunohistochemistry and electron microscopy Many enhancement methods have been reported for DAB involving different buffer conditions the addition of metal ions and post treatment applications Ref 1 7 also see the Pierce Metal Enhanced DAB Substrate Kit Part No 34065 One disadvantage of DAB compared to other substrates is the need to use the working solution immediately because it begins a reaction process upon the addition of hydrogen peroxide This results in increased background color of the substrate However the brown bands resulting from DAB reactions do not fade as quickly as the color products of other precipitating HRP substrates Related Products Pierce DAB Substrate Kit
    Catalog Number:
    34001
    Price:
    None
    Applications:
    Cell Analysis|Cellular Imaging|IHC Staining & Detection|Immunohistochemistry (IHC)|Protein Assays and Analysis|Protein Biology|Western Blotting
    Category:
    Labeling Detection Products
    Buy from Supplier


    Structured Review

    Thermo Fisher dab substrate kit
    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 <t>GFP::mCD8::HRP</t> (J) immunostained with anti-BRP (blue) (I) or labeled with <t>DAB</t> (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.
    Thermo Scientific Pierce DAB enables chromogenic detection of horseradish peroxidase HRP activity based the action of 3 3 diaminobenzidine DAB in Western blot and tissue staining methods DAB 3 3 diaminobenzidine tetrahydrochloride MW 214 1 reacts with HRP in the presence of peroxide to yield an insoluble brown colored product at locations where peroxidase conjugated antibodies are bound to samples The brown precipitate is insoluble in alcohol and other organic solvents making it an excellent substrate for immunohistochemical staining that requires the use of traditional counterstains and mounting media Features of DAB Powder • HRP substrate for detection of horseradish peroxidase activity on solid media including nitrocellulose and PVDF membranes and fixed tissue samples • Chromogenic no special equipment needed for visualization the DAB reaction produces brown bands or spots at sites of reaction with HRP conjugated antibodies or probes • Package options choose powder for custom formulation with buffer and peroxide or a complete kit of preformulated solutions for immediate use DAB has been used in a variety of applications including Western blotting immunohistochemistry and electron microscopy Many enhancement methods have been reported for DAB involving different buffer conditions the addition of metal ions and post treatment applications Ref 1 7 also see the Pierce Metal Enhanced DAB Substrate Kit Part No 34065 One disadvantage of DAB compared to other substrates is the need to use the working solution immediately because it begins a reaction process upon the addition of hydrogen peroxide This results in increased background color of the substrate However the brown bands resulting from DAB reactions do not fade as quickly as the color products of other precipitating HRP substrates Related Products Pierce DAB Substrate Kit
    https://www.bioz.com/result/dab substrate kit/product/Thermo Fisher
    Average 99 stars, based on 95 article reviews
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    dab substrate kit - by Bioz Stars, 2020-11
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    Images

    1) Product Images from "Differing Strategies Despite Shared Lineages of Motor Neurons and Glia to Achieve Robust Development of an Adult Neuropil in Drosophila"

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

    Journal: Neuron

    doi: 10.1016/j.neuron.2018.01.007

    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.
    Figure Legend 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.

    Techniques Used: Marker, Expressing, Labeling, Microscopy

    2) Product Images from "Differing strategies used by motor neurons and glia to achieve robust development of an adult neuropil in Drosophila"

    Article Title: Differing strategies used by motor neurons and glia to achieve robust development of an adult neuropil in Drosophila

    Journal: bioRxiv

    doi: 10.1101/149229

    Cellular organization of the adult thoracic NG. (A-C): Adult VNSs immunostained with anti-BRP (neuropil marker, blue) (A1-C4) and anti-Dll (NG marker, red) (B1-B3) with astrocytes expressing mCD8::GFP (membrane marker, green) (A1-C4), H2B::RFP (Nuclear marker, red) (B1-B3) or the multicolor system FB1.1 (single cell marker, red, yellow and green) (C1-C4) under the control of alrm-Gal4 . (B1-B3) , astrocyte (H2B::RFP+, Dll+): arrow, EG (Dll+): arrowhead. (C1-C4) Different astrocytes with processes occupying different neuropil territories: arrows. See also Figure S1 and video 1. (D-F): Adult VNSs immunostained with anti-BRP (neuropil marker, blue) (D1-F4) and anti-Dll (NG marker, red) ( E1-E3) with EG expressing mCD4::GFP (membrane marker, green) (D1-D3), H2B::RFP (nuclear marker, red) (E1-E3) or the multicolor system FB1.1 (single cell marker, red, yellow and green; ( Hadjieconomou et al., 2011 )) (F1-F4) under the control of R56F03-Gal4 . (E1-E3), astrocyte: arrow (Dll+), EG (H2B::RFP+, Dll+): arrowhead. (F1-F4) Different EG with processes occupying different neuropil territories: arrows. See also Figure S1 and video 1. (G-I): Prothoracic neuromeres with EG expressing mCD4::GFP and immunostained with anti-BRP (blue) and anti-Elav (neuron marker, red) (G) , anti-BRP (blue) and Dll (red) (H) or anti-NCad (neuropil marker, red) and Nrg (axon marker, blue) (I1-2) . (G) Asterisk: Elav+ neuron wrapped by an EG. (H) 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. Enlargements of the boxed regions are to the right of each panel. See also video 2. (J-K): Prothoracic neuromeres with EG expressing mCD4::GFP (J) and GFP::mCD8::HRP (K) immunostained with anti-BRP (blue) (J) or labeled with DAB (Black) (K) . (L) : Low magnification electron microscopy image of the boxed region in (K) . (M-P): enlargement of the boxed regions in (L) . PG: perineurial glia. SPG: subperineurial glia, NG: Neuropil Glia, C: Cortex, Np: Neuropil, NL: neural lamella. (O) : Left, schematic of adult CNS (blue: neuropils, grey: cortex); right, an individual astrocyte and EG labeled with mCD8::GFP under the control of alrm-Gal4 and R56F03-Gal4 using the MARCM technique. Axes: green (posterior), blue (dorsal), red (medial). (R): Average number of NG in the adult VNS, 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 standard deviation. 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.
    Figure Legend Snippet: Cellular organization of the adult thoracic NG. (A-C): Adult VNSs immunostained with anti-BRP (neuropil marker, blue) (A1-C4) and anti-Dll (NG marker, red) (B1-B3) with astrocytes expressing mCD8::GFP (membrane marker, green) (A1-C4), H2B::RFP (Nuclear marker, red) (B1-B3) or the multicolor system FB1.1 (single cell marker, red, yellow and green) (C1-C4) under the control of alrm-Gal4 . (B1-B3) , astrocyte (H2B::RFP+, Dll+): arrow, EG (Dll+): arrowhead. (C1-C4) Different astrocytes with processes occupying different neuropil territories: arrows. See also Figure S1 and video 1. (D-F): Adult VNSs immunostained with anti-BRP (neuropil marker, blue) (D1-F4) and anti-Dll (NG marker, red) ( E1-E3) with EG expressing mCD4::GFP (membrane marker, green) (D1-D3), H2B::RFP (nuclear marker, red) (E1-E3) or the multicolor system FB1.1 (single cell marker, red, yellow and green; ( Hadjieconomou et al., 2011 )) (F1-F4) under the control of R56F03-Gal4 . (E1-E3), astrocyte: arrow (Dll+), EG (H2B::RFP+, Dll+): arrowhead. (F1-F4) Different EG with processes occupying different neuropil territories: arrows. See also Figure S1 and video 1. (G-I): Prothoracic neuromeres with EG expressing mCD4::GFP and immunostained with anti-BRP (blue) and anti-Elav (neuron marker, red) (G) , anti-BRP (blue) and Dll (red) (H) or anti-NCad (neuropil marker, red) and Nrg (axon marker, blue) (I1-2) . (G) Asterisk: Elav+ neuron wrapped by an EG. (H) 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. Enlargements of the boxed regions are to the right of each panel. See also video 2. (J-K): Prothoracic neuromeres with EG expressing mCD4::GFP (J) and GFP::mCD8::HRP (K) immunostained with anti-BRP (blue) (J) or labeled with DAB (Black) (K) . (L) : Low magnification electron microscopy image of the boxed region in (K) . (M-P): enlargement of the boxed regions in (L) . PG: perineurial glia. SPG: subperineurial glia, NG: Neuropil Glia, C: Cortex, Np: Neuropil, NL: neural lamella. (O) : Left, schematic of adult CNS (blue: neuropils, grey: cortex); right, an individual astrocyte and EG labeled with mCD8::GFP under the control of alrm-Gal4 and R56F03-Gal4 using the MARCM technique. Axes: green (posterior), blue (dorsal), red (medial). (R): Average number of NG in the adult VNS, 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 standard deviation. 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.

    Techniques Used: Marker, Expressing, Labeling, Electron Microscopy, Standard Deviation

    3) Product Images from "Immuno- and Enzyme-histochemistry of HRP for Demonstration of Blood Vessel Permeability in Mouse Thymic Tissues by “In Vivo Cryotechnique”"

    Article Title: Immuno- and Enzyme-histochemistry of HRP for Demonstration of Blood Vessel Permeability in Mouse Thymic Tissues by “In Vivo Cryotechnique”

    Journal: Acta Histochemica et Cytochemica

    doi: 10.1267/ahc.14038

    Enzyme-histochemical images of DAB reaction of HRP in cryosections of mouse thymic tissues, which were injected with various concentrations of HRP [10 ( c, d ), 25 ( e, f ), 50 ( g, h ) and 100 mg/ml ( i, j )] via left ventricles, and prepared at 3 min after HRP injection by IVCT. ( a, b ) Control mice (Con) without HRP injection. MG, methyl green staining. ( c–j ) HRP-enzyme reactivities are variously detected in the interstitium (arrows) around some blood vessels in corticomedullary boundary areas and other thick blood vessels of the medulla, depending on the HRP concentration, but they are also found within blood vessels (arrowheads) in addition to tiny blood capillaries. Co, cortex. M, medulla. Bars=20 μm.
    Figure Legend Snippet: Enzyme-histochemical images of DAB reaction of HRP in cryosections of mouse thymic tissues, which were injected with various concentrations of HRP [10 ( c, d ), 25 ( e, f ), 50 ( g, h ) and 100 mg/ml ( i, j )] via left ventricles, and prepared at 3 min after HRP injection by IVCT. ( a, b ) Control mice (Con) without HRP injection. MG, methyl green staining. ( c–j ) HRP-enzyme reactivities are variously detected in the interstitium (arrows) around some blood vessels in corticomedullary boundary areas and other thick blood vessels of the medulla, depending on the HRP concentration, but they are also found within blood vessels (arrowheads) in addition to tiny blood capillaries. Co, cortex. M, medulla. Bars=20 μm.

    Techniques Used: Injection, Mouse Assay, Staining, Concentration Assay

    Enzyme-histochemical images of DAB reaction of HRP in cryosections of mouse thymic tissues, which were injected with HRP (100 mg/ml) via left ventricles, and prepared after various time intervals by IVCT. ( a, b ) Control mice (Con) without HRP injection. Arrowheads: blood vessels. MG, methyl green staining. ( c, d ) At 30 sec, HRP reaction products are slightly detected in the interstitium around some thick blood vessels in corticomedullary boundary areas (arrows) and also within other blood vessels (arrowheads). ( e–h ) At 1 min and 3 min, they are more widely detected in the interstitium around thick blood vessels in corticomedullary boundary areas (arrows) and within some thick blood vessels in both cortical and medullary areas (arrowheads). ( i, j ) At 10 min, they are diffusely detected in all the interstitium of cortical and medullary areas (arrows). ( k, l ) At 15 min, they are uniformly distributed throughout all the interstitium of thymic tissues (arrows) in addition to perivascular spaces of blood vessels (arrowheads). ( m, n ) At 30 min, HRP phagocytosis by macrophages (double arrowheads) is seen to be scattered, which is accompanied by the decrease of DAB reaction intensity in all the interstitial matrix. Arrowhead: blood vessel. Long arrow: nucleus. Short arrow: process. Co, cortex. M, medulla. Bars=20 μm.
    Figure Legend Snippet: Enzyme-histochemical images of DAB reaction of HRP in cryosections of mouse thymic tissues, which were injected with HRP (100 mg/ml) via left ventricles, and prepared after various time intervals by IVCT. ( a, b ) Control mice (Con) without HRP injection. Arrowheads: blood vessels. MG, methyl green staining. ( c, d ) At 30 sec, HRP reaction products are slightly detected in the interstitium around some thick blood vessels in corticomedullary boundary areas (arrows) and also within other blood vessels (arrowheads). ( e–h ) At 1 min and 3 min, they are more widely detected in the interstitium around thick blood vessels in corticomedullary boundary areas (arrows) and within some thick blood vessels in both cortical and medullary areas (arrowheads). ( i, j ) At 10 min, they are diffusely detected in all the interstitium of cortical and medullary areas (arrows). ( k, l ) At 15 min, they are uniformly distributed throughout all the interstitium of thymic tissues (arrows) in addition to perivascular spaces of blood vessels (arrowheads). ( m, n ) At 30 min, HRP phagocytosis by macrophages (double arrowheads) is seen to be scattered, which is accompanied by the decrease of DAB reaction intensity in all the interstitial matrix. Arrowhead: blood vessel. Long arrow: nucleus. Short arrow: process. Co, cortex. M, medulla. Bars=20 μm.

    Techniques Used: Injection, Mouse Assay, Staining, Size-exclusion Chromatography

    Related Articles

    Immunohistochemistry:

    Article Title: Glioblastoma Multiforme Cancer Stem Cells Express Components of the Renin–Angiotensin System
    Article Snippet: .. 3,3-Diaminobenzidine (DAB) IHC staining for SOX2 (1:500; cat# PA094, Thermo Fisher, Scientific, Scoresby, VIC, Australia), PRR (1:2000; cat# ab40790, Abcam, Cambridge, UK), ATIIR1 (1:30; cat# ab9391, Abcam), ATIIR2 (1:2000; cat# NBP1-77368, Novus Biologicals, LLC, Littleton, CO, USA), ACE (1:100; cat# MCA2054, AbD Serotec, Kidlington, UK) diluted with Bond™ primary antibody diluent (cat# AR9352, Leica) was done for all tissue samples. .. Immunofluorescent (IF) IHC staining was performed on two representative GBM tissue samples from the original cohort of patients used for DAB IHC staining, using identical primary antibodies and concentrations.

    Article Title: Vemurafenib Inhibits Active PTK6 in PTEN-null Prostate Tumor Cells
    Article Snippet: .. For immunohistochemistry Ki67 was detected using horseradish peroxidase-conjugated goat anti-rabbit antibodies in the presence of 3,3’-diaminobenzidine (DAB). ..

    Avidin-Biotin Assay:

    Article Title: Preputial gland adenoma in a wild nutria (Myocastor coypus): a case report
    Article Snippet: .. The antigen-antibody complex was labeled with an avidin-biotin peroxidase complex solution (Vector Laboratories, USA) and a DAB substrate kit (Invitrogen, USA). ..

    Labeling:

    Article Title: Preputial gland adenoma in a wild nutria (Myocastor coypus): a case report
    Article Snippet: .. The antigen-antibody complex was labeled with an avidin-biotin peroxidase complex solution (Vector Laboratories, USA) and a DAB substrate kit (Invitrogen, USA). ..

    other:

    Article Title: Isolated oculomotor nerve palsy as a manifestation of diffuse large B cell lymphoma: A case report
    Article Snippet: After applying 3,3′diaminobenzidine (DAB) for color development at room temperature for 5 min, the sections were subsequently counterstained with hematoxylin.

    Expressing:

    Article Title: Differing Strategies Despite Shared Lineages of Motor Neurons and Glia to Achieve Robust Development of an Adult Neuropil in Drosophila
    Article Snippet: .. HRP expression was revealed overnight using the DAB substrate kit (Thermofisher). ..

    Article Title: Differing strategies used by motor neurons and glia to achieve robust development of an adult neuropil in Drosophila
    Article Snippet: .. The expression of the HRP (Horseradish Peroxydase) were revealed overnight using the DAB substrate kit from Thermofisher. ..

    Staining:

    Article Title: Glioblastoma Multiforme Cancer Stem Cells Express Components of the Renin–Angiotensin System
    Article Snippet: .. 3,3-Diaminobenzidine (DAB) IHC staining for SOX2 (1:500; cat# PA094, Thermo Fisher, Scientific, Scoresby, VIC, Australia), PRR (1:2000; cat# ab40790, Abcam, Cambridge, UK), ATIIR1 (1:30; cat# ab9391, Abcam), ATIIR2 (1:2000; cat# NBP1-77368, Novus Biologicals, LLC, Littleton, CO, USA), ACE (1:100; cat# MCA2054, AbD Serotec, Kidlington, UK) diluted with Bond™ primary antibody diluent (cat# AR9352, Leica) was done for all tissue samples. .. Immunofluorescent (IF) IHC staining was performed on two representative GBM tissue samples from the original cohort of patients used for DAB IHC staining, using identical primary antibodies and concentrations.

    Article Title: Severe acute respiratory syndrome coronavirus 3C‐like protease‐induced apoptosis
    Article Snippet: .. The apoptotic cells were further stained using a horseradish peroxidase/diaminobenzidine (HRP/DAB) system of the SuperPicTure™ Polymer Detection Kit (Zymed Laboratories Inc., San Francisco, CA). .. The percentage of apoptotic cells with a permanent intense brown deposit was counted at × 200 magnification using bright field microscopy.

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  • 99
    Thermo Fisher dab substrate kit
    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 <t>GFP::mCD8::HRP</t> (J) immunostained with anti-BRP (blue) (I) or labeled with <t>DAB</t> (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.
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    TH levels in the midbrain of H1+/- mice. No difference in TH level was detected in the VTA between H1+/- and WT mice. (A): Immunofluorescent staining of hnRNP H ( magenta ) and TH ( green ) was conducted in coronal midbrain sections (Bregma: −3.28 mm to −3.64 mm) containing the VTA dopaminergic neurons in adult H1+/- and WT mice. Higher magnification images in panels ( i ) and ( ii ) demonstrate nuclear expression of hnRNP H across all TH-positive dopaminergic neurons that we examined. Scale bars represent 200 μM ( top ) and 20 μM ( bottom) . (B): Representative image showing immunohistochemical <t>DAB</t> staining of TH in coronal sections of the midbrain region (Bregma: −3.28 mm to −3.64 mm). Scale bars represent represent 1 mm. (C-E): Immunohistochemical DAB staining of TH in the midbrain regions revealed no genotypic difference in TH optical density (C) , in number of TH-positive cells (D) , or in the diameter of TH-positive cells (E) between the H1+/- and WT mice [WT, n = 8; H1+/-, n = 8; t(14)
    Pierce Dab Substrate Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (a) Intensity of test line for five independently synthesized batches of PtNCs ( ca . 120 nm) for detection of 50 pg·mL –1 p24 in FBS with 5 min development in <t>CN/DAB</t> and H 2 O 2 . Red line indicates average test line intensity for triplicate measurements across all batches. Coefficient of variation ( n = 5) was 2.4%. (b) Intensity of test line (for detection of 500 pg·mL –1 p24-spiked plasma) against days of test component aging at 44 °C or room temperature (RT). For the aging experiment, nanobody–biotin and either <t>PtNC</t> or HRP–antibody conjugates were lyophilized in appropriate buffers and incubated at specified temperature. At aging time points, spiked plasma was added to the vessel of freeze-dried components and flowed up the LFIA test strip followed by catalytic amplification.
    Pierce Cn Dab, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    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

    Cellular organization of the adult thoracic NG. (A-C): Adult VNSs immunostained with anti-BRP (neuropil marker, blue) (A1-C4) and anti-Dll (NG marker, red) (B1-B3) with astrocytes expressing mCD8::GFP (membrane marker, green) (A1-C4), H2B::RFP (Nuclear marker, red) (B1-B3) or the multicolor system FB1.1 (single cell marker, red, yellow and green) (C1-C4) under the control of alrm-Gal4 . (B1-B3) , astrocyte (H2B::RFP+, Dll+): arrow, EG (Dll+): arrowhead. (C1-C4) Different astrocytes with processes occupying different neuropil territories: arrows. See also Figure S1 and video 1. (D-F): Adult VNSs immunostained with anti-BRP (neuropil marker, blue) (D1-F4) and anti-Dll (NG marker, red) ( E1-E3) with EG expressing mCD4::GFP (membrane marker, green) (D1-D3), H2B::RFP (nuclear marker, red) (E1-E3) or the multicolor system FB1.1 (single cell marker, red, yellow and green; ( Hadjieconomou et al., 2011 )) (F1-F4) under the control of R56F03-Gal4 . (E1-E3), astrocyte: arrow (Dll+), EG (H2B::RFP+, Dll+): arrowhead. (F1-F4) Different EG with processes occupying different neuropil territories: arrows. See also Figure S1 and video 1. (G-I): Prothoracic neuromeres with EG expressing mCD4::GFP and immunostained with anti-BRP (blue) and anti-Elav (neuron marker, red) (G) , anti-BRP (blue) and Dll (red) (H) or anti-NCad (neuropil marker, red) and Nrg (axon marker, blue) (I1-2) . (G) Asterisk: Elav+ neuron wrapped by an EG. (H) 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. Enlargements of the boxed regions are to the right of each panel. See also video 2. (J-K): Prothoracic neuromeres with EG expressing mCD4::GFP (J) and GFP::mCD8::HRP (K) immunostained with anti-BRP (blue) (J) or labeled with DAB (Black) (K) . (L) : Low magnification electron microscopy image of the boxed region in (K) . (M-P): enlargement of the boxed regions in (L) . PG: perineurial glia. SPG: subperineurial glia, NG: Neuropil Glia, C: Cortex, Np: Neuropil, NL: neural lamella. (O) : Left, schematic of adult CNS (blue: neuropils, grey: cortex); right, an individual astrocyte and EG labeled with mCD8::GFP under the control of alrm-Gal4 and R56F03-Gal4 using the MARCM technique. Axes: green (posterior), blue (dorsal), red (medial). (R): Average number of NG in the adult VNS, 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 standard deviation. 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: bioRxiv

    Article Title: Differing strategies used by motor neurons and glia to achieve robust development of an adult neuropil in Drosophila

    doi: 10.1101/149229

    Figure Lengend Snippet: Cellular organization of the adult thoracic NG. (A-C): Adult VNSs immunostained with anti-BRP (neuropil marker, blue) (A1-C4) and anti-Dll (NG marker, red) (B1-B3) with astrocytes expressing mCD8::GFP (membrane marker, green) (A1-C4), H2B::RFP (Nuclear marker, red) (B1-B3) or the multicolor system FB1.1 (single cell marker, red, yellow and green) (C1-C4) under the control of alrm-Gal4 . (B1-B3) , astrocyte (H2B::RFP+, Dll+): arrow, EG (Dll+): arrowhead. (C1-C4) Different astrocytes with processes occupying different neuropil territories: arrows. See also Figure S1 and video 1. (D-F): Adult VNSs immunostained with anti-BRP (neuropil marker, blue) (D1-F4) and anti-Dll (NG marker, red) ( E1-E3) with EG expressing mCD4::GFP (membrane marker, green) (D1-D3), H2B::RFP (nuclear marker, red) (E1-E3) or the multicolor system FB1.1 (single cell marker, red, yellow and green; ( Hadjieconomou et al., 2011 )) (F1-F4) under the control of R56F03-Gal4 . (E1-E3), astrocyte: arrow (Dll+), EG (H2B::RFP+, Dll+): arrowhead. (F1-F4) Different EG with processes occupying different neuropil territories: arrows. See also Figure S1 and video 1. (G-I): Prothoracic neuromeres with EG expressing mCD4::GFP and immunostained with anti-BRP (blue) and anti-Elav (neuron marker, red) (G) , anti-BRP (blue) and Dll (red) (H) or anti-NCad (neuropil marker, red) and Nrg (axon marker, blue) (I1-2) . (G) Asterisk: Elav+ neuron wrapped by an EG. (H) 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. Enlargements of the boxed regions are to the right of each panel. See also video 2. (J-K): Prothoracic neuromeres with EG expressing mCD4::GFP (J) and GFP::mCD8::HRP (K) immunostained with anti-BRP (blue) (J) or labeled with DAB (Black) (K) . (L) : Low magnification electron microscopy image of the boxed region in (K) . (M-P): enlargement of the boxed regions in (L) . PG: perineurial glia. SPG: subperineurial glia, NG: Neuropil Glia, C: Cortex, Np: Neuropil, NL: neural lamella. (O) : Left, schematic of adult CNS (blue: neuropils, grey: cortex); right, an individual astrocyte and EG labeled with mCD8::GFP under the control of alrm-Gal4 and R56F03-Gal4 using the MARCM technique. Axes: green (posterior), blue (dorsal), red (medial). (R): Average number of NG in the adult VNS, 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 standard deviation. 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: The expression of the HRP (Horseradish Peroxydase) were revealed overnight using the DAB substrate kit from Thermofisher.

    Techniques: Marker, Expressing, Labeling, Electron Microscopy, Standard Deviation

    TH levels in the midbrain of H1+/- mice. No difference in TH level was detected in the VTA between H1+/- and WT mice. (A): Immunofluorescent staining of hnRNP H ( magenta ) and TH ( green ) was conducted in coronal midbrain sections (Bregma: −3.28 mm to −3.64 mm) containing the VTA dopaminergic neurons in adult H1+/- and WT mice. Higher magnification images in panels ( i ) and ( ii ) demonstrate nuclear expression of hnRNP H across all TH-positive dopaminergic neurons that we examined. Scale bars represent 200 μM ( top ) and 20 μM ( bottom) . (B): Representative image showing immunohistochemical DAB staining of TH in coronal sections of the midbrain region (Bregma: −3.28 mm to −3.64 mm). Scale bars represent represent 1 mm. (C-E): Immunohistochemical DAB staining of TH in the midbrain regions revealed no genotypic difference in TH optical density (C) , in number of TH-positive cells (D) , or in the diameter of TH-positive cells (E) between the H1+/- and WT mice [WT, n = 8; H1+/-, n = 8; t(14)

    Journal: bioRxiv

    Article Title: A mutation in Hnrnph1 that decreases methamphetamine-induced reinforcement, reward, and dopamine release and increases synaptosomal hnRNP H and mitochondrial proteins

    doi: 10.1101/717728

    Figure Lengend Snippet: TH levels in the midbrain of H1+/- mice. No difference in TH level was detected in the VTA between H1+/- and WT mice. (A): Immunofluorescent staining of hnRNP H ( magenta ) and TH ( green ) was conducted in coronal midbrain sections (Bregma: −3.28 mm to −3.64 mm) containing the VTA dopaminergic neurons in adult H1+/- and WT mice. Higher magnification images in panels ( i ) and ( ii ) demonstrate nuclear expression of hnRNP H across all TH-positive dopaminergic neurons that we examined. Scale bars represent 200 μM ( top ) and 20 μM ( bottom) . (B): Representative image showing immunohistochemical DAB staining of TH in coronal sections of the midbrain region (Bregma: −3.28 mm to −3.64 mm). Scale bars represent represent 1 mm. (C-E): Immunohistochemical DAB staining of TH in the midbrain regions revealed no genotypic difference in TH optical density (C) , in number of TH-positive cells (D) , or in the diameter of TH-positive cells (E) between the H1+/- and WT mice [WT, n = 8; H1+/-, n = 8; t(14)

    Article Snippet: Next, brains were dissected and processed for tyrosine hydroxylase (TH) 3-3’-diaminobenzidine (DAB) IHC and analysis as previously described ( ; ), or double immunofluorescent IHC for hnRNP H and TH colocalization.

    Techniques: Mouse Assay, Staining, Expressing, Immunohistochemistry

    TH levels in striatum of H1+/- mice. No difference in TH level was detected in the striatum between the H1+/- versus WT mice. (A-F): Representative images showing immunohistochemical DAB staining on coronal sections of striatum (Bregma: 1.18 mm to 0.86 mm). A-C: dorsal striatum. D-F: ventral striatum which includes NAc. Scale bars represent represent 1 mm. Optical density (OD) analysis revealed a nonsignificant increase in TH OD in the dorsal striatum [ C: WT, n = 7; H1+/-, n = 8; t(13) = 2.07, p = 0.10; unpaired Student’s T-test] and a small but significant increase in TH intensity in the ventral striatum of H1+/- compared to WT mice [ F : WT, n = 7; H1+/-, n = 8; t(13) = 2.30, *p = 0.04; unpaired Student’s T-test]. (G-H): Immunoblot for TH protein level in the striatum. Representative immunoblot for TH shown in (G) with quantification shown in (H) . There was no change in TH protein expression in the striatum of H1+/- compared to WT mice [WT, n = 14; H1+/-, n = 11; t(23) = 0.62, p = 0.54; unpaired Student’s T-test].

    Journal: bioRxiv

    Article Title: A mutation in Hnrnph1 that decreases methamphetamine-induced reinforcement, reward, and dopamine release and increases synaptosomal hnRNP H and mitochondrial proteins

    doi: 10.1101/717728

    Figure Lengend Snippet: TH levels in striatum of H1+/- mice. No difference in TH level was detected in the striatum between the H1+/- versus WT mice. (A-F): Representative images showing immunohistochemical DAB staining on coronal sections of striatum (Bregma: 1.18 mm to 0.86 mm). A-C: dorsal striatum. D-F: ventral striatum which includes NAc. Scale bars represent represent 1 mm. Optical density (OD) analysis revealed a nonsignificant increase in TH OD in the dorsal striatum [ C: WT, n = 7; H1+/-, n = 8; t(13) = 2.07, p = 0.10; unpaired Student’s T-test] and a small but significant increase in TH intensity in the ventral striatum of H1+/- compared to WT mice [ F : WT, n = 7; H1+/-, n = 8; t(13) = 2.30, *p = 0.04; unpaired Student’s T-test]. (G-H): Immunoblot for TH protein level in the striatum. Representative immunoblot for TH shown in (G) with quantification shown in (H) . There was no change in TH protein expression in the striatum of H1+/- compared to WT mice [WT, n = 14; H1+/-, n = 11; t(23) = 0.62, p = 0.54; unpaired Student’s T-test].

    Article Snippet: Next, brains were dissected and processed for tyrosine hydroxylase (TH) 3-3’-diaminobenzidine (DAB) IHC and analysis as previously described ( ; ), or double immunofluorescent IHC for hnRNP H and TH colocalization.

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Expressing

    (a) Intensity of test line for five independently synthesized batches of PtNCs ( ca . 120 nm) for detection of 50 pg·mL –1 p24 in FBS with 5 min development in CN/DAB and H 2 O 2 . Red line indicates average test line intensity for triplicate measurements across all batches. Coefficient of variation ( n = 5) was 2.4%. (b) Intensity of test line (for detection of 500 pg·mL –1 p24-spiked plasma) against days of test component aging at 44 °C or room temperature (RT). For the aging experiment, nanobody–biotin and either PtNC or HRP–antibody conjugates were lyophilized in appropriate buffers and incubated at specified temperature. At aging time points, spiked plasma was added to the vessel of freeze-dried components and flowed up the LFIA test strip followed by catalytic amplification.

    Journal: ACS Nano

    Article Title: Platinum Nanocatalyst Amplification: Redefining the Gold Standard for Lateral Flow Immunoassays with Ultrabroad Dynamic Range

    doi: 10.1021/acsnano.7b06229

    Figure Lengend Snippet: (a) Intensity of test line for five independently synthesized batches of PtNCs ( ca . 120 nm) for detection of 50 pg·mL –1 p24 in FBS with 5 min development in CN/DAB and H 2 O 2 . Red line indicates average test line intensity for triplicate measurements across all batches. Coefficient of variation ( n = 5) was 2.4%. (b) Intensity of test line (for detection of 500 pg·mL –1 p24-spiked plasma) against days of test component aging at 44 °C or room temperature (RT). For the aging experiment, nanobody–biotin and either PtNC or HRP–antibody conjugates were lyophilized in appropriate buffers and incubated at specified temperature. At aging time points, spiked plasma was added to the vessel of freeze-dried components and flowed up the LFIA test strip followed by catalytic amplification.

    Article Snippet: Next, the strip was immersed in another well for 5 min filled with 330 μL (enough solution to cover test line on strip in well) freshly prepared PtNC development solution containing a modified Pierce CN/DAB (4-chloro-1-naphthol/3,3′-diaminobenzidine, tetrahydrochloride) Substrate Kit (Thermo Scientific) adjusted with hydrogen peroxide solution 30% (w/w) (Sigma) to a final added peroxide concentration of 4 M. Finally, the strip was moved into a well containing 330 μL purified water for 1 min to stop the reaction.

    Techniques: Synthesized, Incubation, Stripping Membranes, Amplification

    (a) Test line intensity obtained by mobile phone camera images for a dilution series of p24 spiked into sera before (black) and after amplification (red) in a mixture of CN/DAB and H 2 O 2 for 5 min using ca . 120 nm PtNC labels. Error bars represent standard deviation from three independent experiments. (b) Illustration of the broad linear dynamic range across 4 orders of magnitude from 10000 to 1 pg·mL –1 achieved with the present method with nonlinear (log scale) regression least-squares fit (predevelopment: 0.62 x + 1.7, R 2 = 0.9813; postdevelopment: 0.61x + 3.0, R 2 = 0.9853). (c) Clinical sample HIV seroconversion panel results for a single donor showing the change in test line intensity as a function of sample blood collection date covering the seroconversion period. 50 μL of each clinical sample was added to a vessel containing lyophilized test reagents and run up the LFIA strip followed by catalytic amplification. Red dotted line indicates cutoff (average test line intensity of 3 nonspiked blank plasma samples plus three times standard deviation). Error bars represent standard deviation from three independent experiments.

    Journal: ACS Nano

    Article Title: Platinum Nanocatalyst Amplification: Redefining the Gold Standard for Lateral Flow Immunoassays with Ultrabroad Dynamic Range

    doi: 10.1021/acsnano.7b06229

    Figure Lengend Snippet: (a) Test line intensity obtained by mobile phone camera images for a dilution series of p24 spiked into sera before (black) and after amplification (red) in a mixture of CN/DAB and H 2 O 2 for 5 min using ca . 120 nm PtNC labels. Error bars represent standard deviation from three independent experiments. (b) Illustration of the broad linear dynamic range across 4 orders of magnitude from 10000 to 1 pg·mL –1 achieved with the present method with nonlinear (log scale) regression least-squares fit (predevelopment: 0.62 x + 1.7, R 2 = 0.9813; postdevelopment: 0.61x + 3.0, R 2 = 0.9853). (c) Clinical sample HIV seroconversion panel results for a single donor showing the change in test line intensity as a function of sample blood collection date covering the seroconversion period. 50 μL of each clinical sample was added to a vessel containing lyophilized test reagents and run up the LFIA strip followed by catalytic amplification. Red dotted line indicates cutoff (average test line intensity of 3 nonspiked blank plasma samples plus three times standard deviation). Error bars represent standard deviation from three independent experiments.

    Article Snippet: Next, the strip was immersed in another well for 5 min filled with 330 μL (enough solution to cover test line on strip in well) freshly prepared PtNC development solution containing a modified Pierce CN/DAB (4-chloro-1-naphthol/3,3′-diaminobenzidine, tetrahydrochloride) Substrate Kit (Thermo Scientific) adjusted with hydrogen peroxide solution 30% (w/w) (Sigma) to a final added peroxide concentration of 4 M. Finally, the strip was moved into a well containing 330 μL purified water for 1 min to stop the reaction.

    Techniques: Amplification, Standard Deviation, Stripping Membranes

    (a) Scheme showing amplified LFIA, where functionalized Pt nanocatalysts (PtNCs) and biotinylated nanobody fragments are mixed with a plasma or serum sample. In the presence of a target, PtNCs become biotinylated through complexation with the target, and rapid high affinity biotin–streptavidin binding enables a [target] dependent deposition of PtNC at the test line. PtNCs bound at the test line catalyze the oxidation of CN/DAB (4-chloro-1-naphthol/3,3′-diaminobenzidine, tetrahydrochloride) substrate in the presence of hydrogen peroxide producing an insoluble black product which is clearly visible with the naked eye. (b) Scheme for site-selective chemical modification of a nanobody with an exposed cysteine mutation (red), where lysine residues are highlighted in orange on the structural model (left), and cartoon of oriented elements at the streptavidin test line. (c) Schematic comparing the dynamic ranges of fourth-generation LFIA, ELISA, and PtNC LFIA.

    Journal: ACS Nano

    Article Title: Platinum Nanocatalyst Amplification: Redefining the Gold Standard for Lateral Flow Immunoassays with Ultrabroad Dynamic Range

    doi: 10.1021/acsnano.7b06229

    Figure Lengend Snippet: (a) Scheme showing amplified LFIA, where functionalized Pt nanocatalysts (PtNCs) and biotinylated nanobody fragments are mixed with a plasma or serum sample. In the presence of a target, PtNCs become biotinylated through complexation with the target, and rapid high affinity biotin–streptavidin binding enables a [target] dependent deposition of PtNC at the test line. PtNCs bound at the test line catalyze the oxidation of CN/DAB (4-chloro-1-naphthol/3,3′-diaminobenzidine, tetrahydrochloride) substrate in the presence of hydrogen peroxide producing an insoluble black product which is clearly visible with the naked eye. (b) Scheme for site-selective chemical modification of a nanobody with an exposed cysteine mutation (red), where lysine residues are highlighted in orange on the structural model (left), and cartoon of oriented elements at the streptavidin test line. (c) Schematic comparing the dynamic ranges of fourth-generation LFIA, ELISA, and PtNC LFIA.

    Article Snippet: Next, the strip was immersed in another well for 5 min filled with 330 μL (enough solution to cover test line on strip in well) freshly prepared PtNC development solution containing a modified Pierce CN/DAB (4-chloro-1-naphthol/3,3′-diaminobenzidine, tetrahydrochloride) Substrate Kit (Thermo Scientific) adjusted with hydrogen peroxide solution 30% (w/w) (Sigma) to a final added peroxide concentration of 4 M. Finally, the strip was moved into a well containing 330 μL purified water for 1 min to stop the reaction.

    Techniques: Amplification, Binding Assay, Modification, Mutagenesis, Enzyme-linked Immunosorbent Assay

    (a) Scheme showing synthesis of Au–Pt core–shell structure (PtNC), where 15 nm gold nanoparticles are used as seeds for subsequent platinum overgrowth in the presence of polyvinylpyrrolidone (PVP) as a stabilizer and l -ascorbic acid as a reducing agent. Influence of PVP molecular weight on PtNC catalytic activity, measured by the absorbance at 652 nm corresponding to the oxidation of TMB by H 2 O 2 ( n = 3). In the absence of PVP, significant aggregation occurred. (b) Catalytic activity of PtNCs incubated in serological and protein-rich environments up to 24 h ( n = 6). (c) Transmission electron micrographs of PtNCs synthesized with varying AuNP seed concentrations to control size: (i) 5 nM seeds and (ii) 0.3 nM seeds. (iii) High-resolution TEM image of an individual PtNC formed from 0.3 nM seed concentration. Inset shows the lattice fringes corresponding to platinum (111) and (200). (iv) Selected area electron diffraction (SAED) pattern taken from a single PtNC ( ca . 120 nm) with diffraction spots consistent with polycrystalline platinum with an FCC lattice. (d) Number distribution of the hydrodynamic diameter of PtNCs formed by varying [Au]:[Pt] measured by dynamic light scattering. Batches were synthesized in the presence of different gold nanoparticle seed concentrations: A, 5 nM; B, 1.2 nM; C, 0.6 nM; D, 0.3 nM; E, 0.15 nM; F, 0.08 nM; G, 75 pM ca . 120 nm PtNCs as seeds; H, 40 pM ca . 120 nm PtNCs as seeds. (e) Intensity of test line for antibody modified PtNCs (150 pM) varying in size from ca . 50 to 280 nm (mean number distribution by DLS) for detection of 100 pg·mL –1 p24 in FBS with 5 min development in CN/DAB and H 2 O 2 . All data are averaged from ≥3 independent measurements where error bars represent the standard deviation from the mean.

    Journal: ACS Nano

    Article Title: Platinum Nanocatalyst Amplification: Redefining the Gold Standard for Lateral Flow Immunoassays with Ultrabroad Dynamic Range

    doi: 10.1021/acsnano.7b06229

    Figure Lengend Snippet: (a) Scheme showing synthesis of Au–Pt core–shell structure (PtNC), where 15 nm gold nanoparticles are used as seeds for subsequent platinum overgrowth in the presence of polyvinylpyrrolidone (PVP) as a stabilizer and l -ascorbic acid as a reducing agent. Influence of PVP molecular weight on PtNC catalytic activity, measured by the absorbance at 652 nm corresponding to the oxidation of TMB by H 2 O 2 ( n = 3). In the absence of PVP, significant aggregation occurred. (b) Catalytic activity of PtNCs incubated in serological and protein-rich environments up to 24 h ( n = 6). (c) Transmission electron micrographs of PtNCs synthesized with varying AuNP seed concentrations to control size: (i) 5 nM seeds and (ii) 0.3 nM seeds. (iii) High-resolution TEM image of an individual PtNC formed from 0.3 nM seed concentration. Inset shows the lattice fringes corresponding to platinum (111) and (200). (iv) Selected area electron diffraction (SAED) pattern taken from a single PtNC ( ca . 120 nm) with diffraction spots consistent with polycrystalline platinum with an FCC lattice. (d) Number distribution of the hydrodynamic diameter of PtNCs formed by varying [Au]:[Pt] measured by dynamic light scattering. Batches were synthesized in the presence of different gold nanoparticle seed concentrations: A, 5 nM; B, 1.2 nM; C, 0.6 nM; D, 0.3 nM; E, 0.15 nM; F, 0.08 nM; G, 75 pM ca . 120 nm PtNCs as seeds; H, 40 pM ca . 120 nm PtNCs as seeds. (e) Intensity of test line for antibody modified PtNCs (150 pM) varying in size from ca . 50 to 280 nm (mean number distribution by DLS) for detection of 100 pg·mL –1 p24 in FBS with 5 min development in CN/DAB and H 2 O 2 . All data are averaged from ≥3 independent measurements where error bars represent the standard deviation from the mean.

    Article Snippet: Next, the strip was immersed in another well for 5 min filled with 330 μL (enough solution to cover test line on strip in well) freshly prepared PtNC development solution containing a modified Pierce CN/DAB (4-chloro-1-naphthol/3,3′-diaminobenzidine, tetrahydrochloride) Substrate Kit (Thermo Scientific) adjusted with hydrogen peroxide solution 30% (w/w) (Sigma) to a final added peroxide concentration of 4 M. Finally, the strip was moved into a well containing 330 μL purified water for 1 min to stop the reaction.

    Techniques: Molecular Weight, Activity Assay, Incubation, Transmission Assay, Synthesized, Transmission Electron Microscopy, Concentration Assay, Modification, Standard Deviation