vectastain  (Vector Laboratories)


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    VECTASTAIN ABC AmP Reagent for Western Blot Detection Standard Kit
    Description:
    VECTASTAIN AMP Amplifed Detection Greater sensitivity can be achieved by using an amplified detection procedure Our biotin avidin and biotin streptavidin systems introduce a large number of enzymes to the target site providing signal amplification while maintaining low background The VECTASTAIN ABC AmP is an amplified ABC alkaline phosphatase reagent for the detection of mouse or rabbit primary antibodies on nitrocellulose or PVDF membranes The signal can be visualized using either a chemiluminescent chemifluorescent or a chromogenic substrate Used with the DuoLuX Chemiluminescent Chemifluorescent Substrate the VECTASTAIN ABC AmP system produces a very high and sustained light emission signal with low background and permanent fluorescence The sensitivity of VECTASTAIN ABC AmP Reagent combined with the DuoLux substrate allows for the detection of as little as 1 pg of target protein The VECTASTAIN ABC AmP Reagent is sufficient to develop approximately twenty 100 cm2blots
    Catalog Number:
    AK-6000
    Price:
    None
    Category:
    Protein chromogenic detection reagents or kits or substrates
    Reactivity:
    No antibody included
    Size:
    1 Kit
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    Structured Review

    Vector Laboratories vectastain
    VECTASTAIN ABC AmP Reagent for Western Blot Detection Standard Kit
    VECTASTAIN AMP Amplifed Detection Greater sensitivity can be achieved by using an amplified detection procedure Our biotin avidin and biotin streptavidin systems introduce a large number of enzymes to the target site providing signal amplification while maintaining low background The VECTASTAIN ABC AmP is an amplified ABC alkaline phosphatase reagent for the detection of mouse or rabbit primary antibodies on nitrocellulose or PVDF membranes The signal can be visualized using either a chemiluminescent chemifluorescent or a chromogenic substrate Used with the DuoLuX Chemiluminescent Chemifluorescent Substrate the VECTASTAIN ABC AmP system produces a very high and sustained light emission signal with low background and permanent fluorescence The sensitivity of VECTASTAIN ABC AmP Reagent combined with the DuoLux substrate allows for the detection of as little as 1 pg of target protein The VECTASTAIN ABC AmP Reagent is sufficient to develop approximately twenty 100 cm2blots
    https://www.bioz.com/result/vectastain/product/Vector Laboratories
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vectastain - by Bioz Stars, 2021-04
    98/100 stars

    Images

    1) Product Images from "Mitochondrial transplantation enhances murine lung viability and recovery after ischemia-reperfusion injury"

    Article Title: Mitochondrial transplantation enhances murine lung viability and recovery after ischemia-reperfusion injury

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00221.2019

    Mitochondrial biodistribution and tissue uptake. PET-microcomputed tomography imaging of 18 F-labeled rhodamine 6G-labeled mitochondria (1 × 10 9 , red) delivered to the lungs of Wistar male rats ( n = 6; A and C ). A : in rats receiving mitochondria via the pulmonary artery ( n = 3), 18 F-labeled rhodamine 6G mitochondria (1 × 10 9 mitochondria in 0.5 mL buffer) were injected to the lungs as a bolus antegrade via the pulmonary trunk using a tuberculin syringe with a 30-gauge needle. C : in rats receiving mitochondria via nebulization ( n = 3), 18 F-labeled rhodamine 6G labeled mitochondria (1 × 10 9 in 0.3 mL buffer) were delivered as an aerosol to the lungs via the trachea via nebulization using the Aeroneb ultrasonic nebulizer connected to the FlexVent system. Wistar rats were allowed to recover for 10 min after mitochondrial delivery and were then euthanized in a CO 2 chamber before imaging. Labeled mitochondria (red signal on the imaging) were detected only in the lungs; radiolabeled mitochondria were not detectable in any other organ or region of the body ( A and C ). Representative immunohistochemical images (×100) of human mitochondria delivered to the lungs of C57BL/6J male mice at the beginning of reperfusion following 2 h of ischemia ( n = 6; B and D ). B : in mice receiving human mitochondria via the pulmonary artery ( n = 3), human mitochondria (1 × 10 8 mitochondria in 0.3 mL buffer) were injected to the lung as a bolus antegrade via the left pulmonary artery using a tuberculin syringe with a 40-gauge needle. D : in mice receiving human mitochondria via nebulization ( n = 3), human mitochondria (3 × 10 8 in 0.3 mL buffer) were delivered as an aerosol to the lungs via the trachea via nebulization using the Aeroneb ultrasonic nebulizer connected to the FlexVent system. C57BL/6J mice were allowed to recover for 30 min after mitochondrial delivery. Left lung tissue was then harvested for further immnohistological staining. Human mitochondria were detected using monoclonal anti-human mitochondrial antibody (biotin-MTCO2, Abcam, Cambridge, MA) and visualized using Vectastain (Vector Laboratories, Burlingame, CA) and AEC+ substrate chromogen (Dako, Agilent, Santa Clara, CA). Human mitochondria indicated by black arrows were globally distributed throughout the lung and detected within and around lung alveoli (a) and connective tissue ( B and D ). μCT, microcomputed tomography.
    Figure Legend Snippet: Mitochondrial biodistribution and tissue uptake. PET-microcomputed tomography imaging of 18 F-labeled rhodamine 6G-labeled mitochondria (1 × 10 9 , red) delivered to the lungs of Wistar male rats ( n = 6; A and C ). A : in rats receiving mitochondria via the pulmonary artery ( n = 3), 18 F-labeled rhodamine 6G mitochondria (1 × 10 9 mitochondria in 0.5 mL buffer) were injected to the lungs as a bolus antegrade via the pulmonary trunk using a tuberculin syringe with a 30-gauge needle. C : in rats receiving mitochondria via nebulization ( n = 3), 18 F-labeled rhodamine 6G labeled mitochondria (1 × 10 9 in 0.3 mL buffer) were delivered as an aerosol to the lungs via the trachea via nebulization using the Aeroneb ultrasonic nebulizer connected to the FlexVent system. Wistar rats were allowed to recover for 10 min after mitochondrial delivery and were then euthanized in a CO 2 chamber before imaging. Labeled mitochondria (red signal on the imaging) were detected only in the lungs; radiolabeled mitochondria were not detectable in any other organ or region of the body ( A and C ). Representative immunohistochemical images (×100) of human mitochondria delivered to the lungs of C57BL/6J male mice at the beginning of reperfusion following 2 h of ischemia ( n = 6; B and D ). B : in mice receiving human mitochondria via the pulmonary artery ( n = 3), human mitochondria (1 × 10 8 mitochondria in 0.3 mL buffer) were injected to the lung as a bolus antegrade via the left pulmonary artery using a tuberculin syringe with a 40-gauge needle. D : in mice receiving human mitochondria via nebulization ( n = 3), human mitochondria (3 × 10 8 in 0.3 mL buffer) were delivered as an aerosol to the lungs via the trachea via nebulization using the Aeroneb ultrasonic nebulizer connected to the FlexVent system. C57BL/6J mice were allowed to recover for 30 min after mitochondrial delivery. Left lung tissue was then harvested for further immnohistological staining. Human mitochondria were detected using monoclonal anti-human mitochondrial antibody (biotin-MTCO2, Abcam, Cambridge, MA) and visualized using Vectastain (Vector Laboratories, Burlingame, CA) and AEC+ substrate chromogen (Dako, Agilent, Santa Clara, CA). Human mitochondria indicated by black arrows were globally distributed throughout the lung and detected within and around lung alveoli (a) and connective tissue ( B and D ). μCT, microcomputed tomography.

    Techniques Used: Positron Emission Tomography, Imaging, Labeling, Injection, Immunohistochemistry, Mouse Assay, Staining, Plasmid Preparation

    Related Articles

    Binding Assay:

    Article Title: Low doses of bioherbicide favour prion aggregation and propagation in vivo
    Article Snippet: .. The SAF84 antibody was used to label PrPSc and the Vectastain ABC-AmP kit (Vector laboratories, USA) to reveal antibody binding. .. Softwares and statistical analyses Kaplan-Meier survival curves were done using the GraphPad Prism software (La Jolla, CA, USA).

    Article Title: A Fluorescent Oligothiophene-Bis-Triazine ligand interacts with PrP fibrils and detects SDS-resistant oligomers in human prion diseases
    Article Snippet: .. The SAF84 antibody was used to label rPrPSc and the Vectastain ABC-AmP kit (Vector laboratories, USA) to reveal antibody binding. .. Software and statistical analysis ImageJ ( http://rsb.info.nih.gov/ij/index.html ) was used to compare the intensity of individual western blot bands.

    Article Title: Plasma cholesterol level determines in vivo prion propagation [S]
    Article Snippet: .. The SAF84 antibody was used to label PrPSc and the Vectastain ABC-AmP kit (Vector Laboratories) was used to reveal antibody binding. ..

    FLAG-tag:

    Article Title: Selective incorporation of influenza virus RNA segments into virions
    Article Snippet: .. Antigens were detected with anti-FLAG monoclonal antibody M2 (Sigma) or rabbit antiserum against influenza WSN virus used as the primary antibody and biotinylated anti-mouse IgG for the FLAG epitope or biotinylated anti-rabbit IgG for viral antigens used as the secondary antibody (VECTASTAIN ABC kit, Vector Laboratories). .. Cells were fixed with 3% formaldehyde solution, permeated by 0.1% Triton X-100 in 3% formaldehyde solution, and prehybridized at 65°C for 30 min in prehybridization buffer [5× SSC (1× SSC = 0.15 M sodium chloride/0.015 M sodium citrate, pH 7.0)/1% blocking reagent/DIG nucleic acid-detection kit (Roche Diagnostics, Indianapolis)/0.1% N -lauroylsarcosine/0.02% SDS containing 0.1 mg/ml poly(A) DNA from the DIG-detection kit)].

    Incubation:

    Article Title: Immunohistochemical analysis of macroautophagy
    Article Snippet: Subsequently, the primary antibody was applied overnight in a damp box, followed by 2 × 5 min immersion in staining buffer. .. Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer. .. In case the primary antibody was already biotinylated, blocking with normal serum and incubation with biotinylated secondary antibodies were omitted.

    Article Title: The Hemagglutinin-Neuraminidase Protein of Newcastle Disease Virus Determines Tropism and Virulence
    Article Snippet: The sections were then incubated with a 1:500 dilution of the primary NDV monoclonal antibody ( ) cocktail against HN (10D11, AVS, 15C4, and B79) for 1 h at room temperature. .. After a wash, sections were incubated with the VectaStain secondary antibody (Vector Laboratories) for 30 min, as recommended by the manufacturer. .. After a further wash cycle, the sections were incubated with the VectaStain ABC reagent for 30 min.

    Staining:

    Article Title: Immunohistochemical analysis of macroautophagy
    Article Snippet: Subsequently, the primary antibody was applied overnight in a damp box, followed by 2 × 5 min immersion in staining buffer. .. Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer. .. In case the primary antibody was already biotinylated, blocking with normal serum and incubation with biotinylated secondary antibodies were omitted.

    Immunostaining:

    Article Title: Involvement of TRPC Channels in Lung Cancer Cell Differentiation and the Correlation Analysis in Human Non-Small Cell Lung Cancer
    Article Snippet: .. The immunostaining procedure was similar to our reports , and the VECTASTAIN ABC system (Vector Laboratories, Peterborough, UK) was used. .. The rabbit anti-TRPC1, 3, 4 and 6 antibodies purchased from Abcam (Cambridge, UK) were used for human lung tissue and lung cancer section staining.

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    • vectastain abc kit  (Vector Laboratories)
    98
    Vector Laboratories vectastain abc kit
    Confirmation of virus production with truncated NA RNA segments. ( A ) MDCK cells were infected with NA(−) ( a and d ), NAFLAG ( b and e ), or NAFLAGMet(−) ( c and f ) viruses and overlaid with 0.6% agarose. After incubation for 48 h at 37°C, the cells were fixed and permeated with 0.1% Triton X-100 in 3% formaldehyde solution. The viral proteins or FLAG epitope were detected by immunostaining with antiserum to influenza WSN strain ( a – c ) or anti-FLAG monoclonal antibody ( d – f ) as the primary antibody and biotinylated secondary antibody with the <t>VECTASTAIN</t> <t>ABC</t> kit (Vector Laboratories). ( B ) MDCK cells infected with NA(−), NAFLAG, or NAFLAGMet(−) viruses were incubated, fixed, and permeated as described above. The FLAG sequence in mRNA was detected by in situ hybridization with a digoxigenin-labeled probe specific for the sequence and visualized with the DIG nucleic acid-detection kit (Roche).
    Vectastain Abc Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vectastain abc kit/product/Vector Laboratories
    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vectastain abc kit - by Bioz Stars, 2021-04
    98/100 stars
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    Image Search Results


    Confirmation of virus production with truncated NA RNA segments. ( A ) MDCK cells were infected with NA(−) ( a and d ), NAFLAG ( b and e ), or NAFLAGMet(−) ( c and f ) viruses and overlaid with 0.6% agarose. After incubation for 48 h at 37°C, the cells were fixed and permeated with 0.1% Triton X-100 in 3% formaldehyde solution. The viral proteins or FLAG epitope were detected by immunostaining with antiserum to influenza WSN strain ( a – c ) or anti-FLAG monoclonal antibody ( d – f ) as the primary antibody and biotinylated secondary antibody with the VECTASTAIN ABC kit (Vector Laboratories). ( B ) MDCK cells infected with NA(−), NAFLAG, or NAFLAGMet(−) viruses were incubated, fixed, and permeated as described above. The FLAG sequence in mRNA was detected by in situ hybridization with a digoxigenin-labeled probe specific for the sequence and visualized with the DIG nucleic acid-detection kit (Roche).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Selective incorporation of influenza virus RNA segments into virions

    doi: 10.1073/pnas.0437772100

    Figure Lengend Snippet: Confirmation of virus production with truncated NA RNA segments. ( A ) MDCK cells were infected with NA(−) ( a and d ), NAFLAG ( b and e ), or NAFLAGMet(−) ( c and f ) viruses and overlaid with 0.6% agarose. After incubation for 48 h at 37°C, the cells were fixed and permeated with 0.1% Triton X-100 in 3% formaldehyde solution. The viral proteins or FLAG epitope were detected by immunostaining with antiserum to influenza WSN strain ( a – c ) or anti-FLAG monoclonal antibody ( d – f ) as the primary antibody and biotinylated secondary antibody with the VECTASTAIN ABC kit (Vector Laboratories). ( B ) MDCK cells infected with NA(−), NAFLAG, or NAFLAGMet(−) viruses were incubated, fixed, and permeated as described above. The FLAG sequence in mRNA was detected by in situ hybridization with a digoxigenin-labeled probe specific for the sequence and visualized with the DIG nucleic acid-detection kit (Roche).

    Article Snippet: Antigens were detected with anti-FLAG monoclonal antibody M2 (Sigma) or rabbit antiserum against influenza WSN virus used as the primary antibody and biotinylated anti-mouse IgG for the FLAG epitope or biotinylated anti-rabbit IgG for viral antigens used as the secondary antibody (VECTASTAIN ABC kit, Vector Laboratories).

    Techniques: Infection, Incubation, FLAG-tag, Immunostaining, Plasmid Preparation, Sequencing, In Situ Hybridization, Labeling

    Figure 6. Immunohistochemical detection of LC3B is feasible in liver from starved Gfp-Lc3 tg/tg mice using Vectastain ABC. ( A ) Western blot analysis of LC3B in liver from wild-type mice, heterozygous Gfp-Lc3 tg/+ or homozygous Gfp-Lc3 tg/tg transgenic mice. The GFP-LC3 expression in each group was quantified. **p

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 6. Immunohistochemical detection of LC3B is feasible in liver from starved Gfp-Lc3 tg/tg mice using Vectastain ABC. ( A ) Western blot analysis of LC3B in liver from wild-type mice, heterozygous Gfp-Lc3 tg/+ or homozygous Gfp-Lc3 tg/tg transgenic mice. The GFP-LC3 expression in each group was quantified. **p

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Immunohistochemistry, Mouse Assay, Western Blot, Transgenic Assay, Expressing

    Figure 15. SQSTM1 accumulates in cytoplasmic inclusions of liver from autophagy-deficient mice. Liver samples were isolated from fed Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for SQSTM1 using rabbit polyclonal anti-SQSTM1 (Sigma, 1:5,000) and Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm. The positive area of SQSTM1 inclusions was quantified. ***p

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 15. SQSTM1 accumulates in cytoplasmic inclusions of liver from autophagy-deficient mice. Liver samples were isolated from fed Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for SQSTM1 using rabbit polyclonal anti-SQSTM1 (Sigma, 1:5,000) and Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm. The positive area of SQSTM1 inclusions was quantified. ***p

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Mouse Assay, Isolation, Staining

    Figure 5. The highest expression of LC3A and LC3B in nonstarved control mice was found in brain tissue. ( A ) Western blot analysis of LC3A and LC3B in different tissue lysates. GAPDH served as a loading control. ( B ) Immunohistochemical detection of LC3A and LC3B in mouse brain using the Vectastain ABC system. Tissue samples were isolated from fed control mice. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for LC3A ( A ) and LC3B ( B ) using rabbit polyclonal anti-LC3A (Abgent, 1:3000) and biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100). Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm.

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 5. The highest expression of LC3A and LC3B in nonstarved control mice was found in brain tissue. ( A ) Western blot analysis of LC3A and LC3B in different tissue lysates. GAPDH served as a loading control. ( B ) Immunohistochemical detection of LC3A and LC3B in mouse brain using the Vectastain ABC system. Tissue samples were isolated from fed control mice. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for LC3A ( A ) and LC3B ( B ) using rabbit polyclonal anti-LC3A (Abgent, 1:3000) and biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100). Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm.

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Expressing, Mouse Assay, Western Blot, Immunohistochemistry, Isolation, Staining

    Figure 13. ATG5 and CTSD are not suitable targets for the immunohistochemical detection of autophagy in liver. Liver samples were isolated from fed Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for ATG5 ( A ) or CTSD ( B ) using rabbit polyclonal anti-ATG5 (Abcam, 1:100) and rabbit monoclonal anti-CTSD (clone EPR3057Y, Abcam, 1:1000), respectively, in combination with Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm. The ATG5- and CTSD-positive area was quantified. Neither the effect of starvation nor the results between Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice were statistically significant.

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 13. ATG5 and CTSD are not suitable targets for the immunohistochemical detection of autophagy in liver. Liver samples were isolated from fed Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for ATG5 ( A ) or CTSD ( B ) using rabbit polyclonal anti-ATG5 (Abcam, 1:100) and rabbit monoclonal anti-CTSD (clone EPR3057Y, Abcam, 1:1000), respectively, in combination with Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm. The ATG5- and CTSD-positive area was quantified. Neither the effect of starvation nor the results between Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice were statistically significant.

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Immunohistochemistry, Isolation, Mouse Assay, Staining

    Figure 7. Optimal immunohistochemical detection of LC3B requires processing of tissue samples in a suitable fixative. Liver samples were isolated from Gfp-Lc3 tg/tg mice that underwent starvation for 48 h. After fixation in different fixatives for 24 h, tissues were paraffin-embedded and stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100) and Vectastain ABC. For formalin-fixed samples, heat-mediated antigen retrieval was performed either in citrate buffer (pH 6.0) or in EDTA buffer (pH 8.0). Scale bar, 20 μm.

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 7. Optimal immunohistochemical detection of LC3B requires processing of tissue samples in a suitable fixative. Liver samples were isolated from Gfp-Lc3 tg/tg mice that underwent starvation for 48 h. After fixation in different fixatives for 24 h, tissues were paraffin-embedded and stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100) and Vectastain ABC. For formalin-fixed samples, heat-mediated antigen retrieval was performed either in citrate buffer (pH 6.0) or in EDTA buffer (pH 8.0). Scale bar, 20 μm.

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Immunohistochemistry, Isolation, Mouse Assay, Staining

    Figure 4. Liver from autophagy-deficient Atg7 F/F Alb-Cre + mice but not from autophagy-competent Atg7 +/+ Alb-Cre + shows immunohistochemical staining for LC3A and LC3B. Liver samples were isolated from fed mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for LC3A ( A ) and LC3B ( B ) using rabbit polyclonal anti-LC3A (Abgent, 1:100) and biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100) in combination with Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm. The LC3A and LC3B positive area was quantified. ***p

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 4. Liver from autophagy-deficient Atg7 F/F Alb-Cre + mice but not from autophagy-competent Atg7 +/+ Alb-Cre + shows immunohistochemical staining for LC3A and LC3B. Liver samples were isolated from fed mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for LC3A ( A ) and LC3B ( B ) using rabbit polyclonal anti-LC3A (Abgent, 1:100) and biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100) in combination with Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm. The LC3A and LC3B positive area was quantified. ***p

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Isolation

    Figure 9. LC3B dots are detectable in frozen liver sections using a staining procedure with alkaline phosphatase and are localized on the surface of lipid droplets. Liver samples were isolated from Atg7 +/+ Alb-Cre + ( A ) or Atg7 F/F Alb-Cre + mice ( B ) that were fed normal chow (control) or underwent starvation for 48 h. After fixation in 4% paraformaldehyde, frozen sections were stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:1,000) and Vectastain ABC-AP kit, containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase. LC3B-positive dots (arrows) were detected on the surface of lipid droplets. These structures could be stained using oil red O. Scale bar, 20 μm.

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 9. LC3B dots are detectable in frozen liver sections using a staining procedure with alkaline phosphatase and are localized on the surface of lipid droplets. Liver samples were isolated from Atg7 +/+ Alb-Cre + ( A ) or Atg7 F/F Alb-Cre + mice ( B ) that were fed normal chow (control) or underwent starvation for 48 h. After fixation in 4% paraformaldehyde, frozen sections were stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:1,000) and Vectastain ABC-AP kit, containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase. LC3B-positive dots (arrows) were detected on the surface of lipid droplets. These structures could be stained using oil red O. Scale bar, 20 μm.

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Staining, Isolation, Mouse Assay

    Figure 8. Immunohistochemical staining of LC3B is enhanced in frozen liver sections. Liver samples were isolated from Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice ( A ) or from transgenic Gfp-Lc3 tg/+ or Gfp-Lc3 tg/tg mice ( B ). Some animals were fed normal chow (control), others underwent starvation for 48 h. After fixation in acetone, frozen sections were stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:1,000 [ Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + samples] or 1:30,000 [ Gfp-Lc3 tg/+ and Gfp-Lc3 tg/tg samples]) and Vectastain ABC. Scale bar, 40 μm.

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 8. Immunohistochemical staining of LC3B is enhanced in frozen liver sections. Liver samples were isolated from Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice ( A ) or from transgenic Gfp-Lc3 tg/+ or Gfp-Lc3 tg/tg mice ( B ). Some animals were fed normal chow (control), others underwent starvation for 48 h. After fixation in acetone, frozen sections were stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:1,000 [ Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + samples] or 1:30,000 [ Gfp-Lc3 tg/+ and Gfp-Lc3 tg/tg samples]) and Vectastain ABC. Scale bar, 40 μm.

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Immunohistochemistry, Staining, Isolation, Mouse Assay, Transgenic Assay

    Figure 14. BECN1 is upregulated in mouse tissue after starvation. ( A ) Liver samples were isolated from fed Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for BECN1 using rabbit polyclonal anti-BECN1 (Lifespan, 1:1,000) and Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar 20 μm. The BECN1 positive area was quantified. ***p

    Journal: Autophagy

    Article Title: Immunohistochemical analysis of macroautophagy

    doi: 10.4161/auto.22968

    Figure Lengend Snippet: Figure 14. BECN1 is upregulated in mouse tissue after starvation. ( A ) Liver samples were isolated from fed Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for BECN1 using rabbit polyclonal anti-BECN1 (Lifespan, 1:1,000) and Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar 20 μm. The BECN1 positive area was quantified. ***p

    Article Snippet: Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer.

    Techniques: Isolation, Mouse Assay, Staining

    The PrP Sc amyloid load is substantially decreased in PLTP −/− mice fed a standard chow diet. A, B: PET blot analyses of frontal tissue sections of WT or PLTP −/− mice not inoculated with prions and euthanized at 233 days (dpi) while they were healthy. C–E: PET blot analyses of frontal tissue sections of WT or PLTP −/− mice inoculated with 22L prion strain, fed a standard chow diet or Western-type cholesterol-rich diet, and all euthanized on the same day (196 dpi), when they were in an asymptomatic stage (C, D) or while they were at the terminal stage of the disease (E). The SAF84 antibody was used to detect PrP SC proteins and the Vectastain ABC-AmP kit (Vector Laboratories) was used to reveal antibody binding.

    Journal: Journal of Lipid Research

    Article Title: Plasma cholesterol level determines in vivo prion propagation [S]

    doi: 10.1194/jlr.M073718

    Figure Lengend Snippet: The PrP Sc amyloid load is substantially decreased in PLTP −/− mice fed a standard chow diet. A, B: PET blot analyses of frontal tissue sections of WT or PLTP −/− mice not inoculated with prions and euthanized at 233 days (dpi) while they were healthy. C–E: PET blot analyses of frontal tissue sections of WT or PLTP −/− mice inoculated with 22L prion strain, fed a standard chow diet or Western-type cholesterol-rich diet, and all euthanized on the same day (196 dpi), when they were in an asymptomatic stage (C, D) or while they were at the terminal stage of the disease (E). The SAF84 antibody was used to detect PrP SC proteins and the Vectastain ABC-AmP kit (Vector Laboratories) was used to reveal antibody binding.

    Article Snippet: The SAF84 antibody was used to label PrPSc and the Vectastain ABC-AmP kit (Vector Laboratories) was used to reveal antibody binding.

    Techniques: Mouse Assay, Positron Emission Tomography, Western Blot, Plasmid Preparation, Binding Assay

    Pre-incubation of brain inoculum with MR100 substantially increases the survival time of injected animals. a Table with the number of animals sacrificed with symptoms before 315 days post-inoculation compared to the total number of animals for each group. b Kaplan-Meier survival plots of mice inoculated (15 μL/each mouse) with a 22 L-infected brain homogenate that was previously diluted in 2 % Sarkosyl/PBS and incubated at a final concentration of 1.5 mM MR100 (n = 8; 22 L + MR100; red) or an equivalent volume of DMSO (150 μL) (n = 10; 22 L + DM; blue) or PBS (150 μL) (n = 10; 22 L+ PBS; black) at room temperature for 2 h. c Histological analyses of thalamus (Th) sections from mice not inoculated with prions (1), or that were inoculated with 22 L + DM inoculum (2) or 22 L + MR100 inoculum and presented no symptoms (3) or with symptoms (4) of prion disease when they were killed (d.p.i., days post-inoculation). Tissue sections were stained with hematoxylin and eosin (HE) to confirm the prion pathology as indicated by the presence of vacuoles and probed with anti-GFAP antibodies as a marker of astrocytic gliosis. Tissue labeling was performed on several animals (2-3 per group) and images are representative of the staining observed in each group. d PET-blot analysis of frontal tissue sections of mice non-inoculated with prions (1), or that were inoculated with 22 L + DM inoculum (2), or 22 L + MR100 inoculum and presented no symptoms (3) or with symptoms (4) of prion disease when they were killed. The SAF84 antibody was used to detect rPrP Sc proteins and the Vectastain ABC-AmP Kit (Vector laboratories, USA) to reveal antibody binding. Scale bars, 10 μM, w: with, w.o.: without symptoms

    Journal: Molecular Neurodegeneration

    Article Title: A Fluorescent Oligothiophene-Bis-Triazine ligand interacts with PrP fibrils and detects SDS-resistant oligomers in human prion diseases

    doi: 10.1186/s13024-016-0074-7

    Figure Lengend Snippet: Pre-incubation of brain inoculum with MR100 substantially increases the survival time of injected animals. a Table with the number of animals sacrificed with symptoms before 315 days post-inoculation compared to the total number of animals for each group. b Kaplan-Meier survival plots of mice inoculated (15 μL/each mouse) with a 22 L-infected brain homogenate that was previously diluted in 2 % Sarkosyl/PBS and incubated at a final concentration of 1.5 mM MR100 (n = 8; 22 L + MR100; red) or an equivalent volume of DMSO (150 μL) (n = 10; 22 L + DM; blue) or PBS (150 μL) (n = 10; 22 L+ PBS; black) at room temperature for 2 h. c Histological analyses of thalamus (Th) sections from mice not inoculated with prions (1), or that were inoculated with 22 L + DM inoculum (2) or 22 L + MR100 inoculum and presented no symptoms (3) or with symptoms (4) of prion disease when they were killed (d.p.i., days post-inoculation). Tissue sections were stained with hematoxylin and eosin (HE) to confirm the prion pathology as indicated by the presence of vacuoles and probed with anti-GFAP antibodies as a marker of astrocytic gliosis. Tissue labeling was performed on several animals (2-3 per group) and images are representative of the staining observed in each group. d PET-blot analysis of frontal tissue sections of mice non-inoculated with prions (1), or that were inoculated with 22 L + DM inoculum (2), or 22 L + MR100 inoculum and presented no symptoms (3) or with symptoms (4) of prion disease when they were killed. The SAF84 antibody was used to detect rPrP Sc proteins and the Vectastain ABC-AmP Kit (Vector laboratories, USA) to reveal antibody binding. Scale bars, 10 μM, w: with, w.o.: without symptoms

    Article Snippet: The SAF84 antibody was used to label rPrPSc and the Vectastain ABC-AmP kit (Vector laboratories, USA) to reveal antibody binding.

    Techniques: Incubation, Injection, Mouse Assay, Infection, Concentration Assay, Staining, Marker, Labeling, Positron Emission Tomography, Plasmid Preparation, Binding Assay