vectastain abc reagent  (Vector Laboratories)


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    Name:
    VECTASTAIN ABC AP Staining KIT Alkaline Phosphatase Standard
    Description:
    VECTASTAIN ABC AP Staining Kits remains widely popular with more than 50 000 citations to its credit Based on the versatile avidin biotin complex interaction the system is modular and along with our selection of secondary antibodies can accommodate a wide array of primary antibody and tissue species Our ABC kits are economical and continue to be a staple product in any immunohistochemistry IHC and immunocytochemistry ICC laboratory The sensitivity of the VECTASTAIN ABC AP system is comparable to that of the peroxidase VECTASTAIN Elite ABC system The VECTASTAIN ABC AP Kits may be preferred for tissues that have high endogenous peroxidase activity The system also offers additional substrate color choices VECTASTAIN ABC AP Kit Components 2 ml Reagent A2 ml Reagent BReagent A and B when mixed form the ABC AP complex Kit is sufficient to stain approximately 1000 2000 sections or fifty 100 cm2 blots
    Catalog Number:
    AK-5000
    Price:
    None
    Category:
    Protein chromogenic detection reagents or kits or substrates
    Reactivity:
    No antibody included
    Size:
    1 Kit
    Buy from Supplier


    Structured Review

    Vector Laboratories vectastain abc reagent
    VECTASTAIN ABC AP Staining KIT Alkaline Phosphatase Standard
    VECTASTAIN ABC AP Staining Kits remains widely popular with more than 50 000 citations to its credit Based on the versatile avidin biotin complex interaction the system is modular and along with our selection of secondary antibodies can accommodate a wide array of primary antibody and tissue species Our ABC kits are economical and continue to be a staple product in any immunohistochemistry IHC and immunocytochemistry ICC laboratory The sensitivity of the VECTASTAIN ABC AP system is comparable to that of the peroxidase VECTASTAIN Elite ABC system The VECTASTAIN ABC AP Kits may be preferred for tissues that have high endogenous peroxidase activity The system also offers additional substrate color choices VECTASTAIN ABC AP Kit Components 2 ml Reagent A2 ml Reagent BReagent A and B when mixed form the ABC AP complex Kit is sufficient to stain approximately 1000 2000 sections or fifty 100 cm2 blots
    https://www.bioz.com/result/vectastain abc reagent/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vectastain abc reagent - by Bioz Stars, 2021-09
    99/100 stars

    Images

    1) Product Images from "Upregulation of GRAIL Is Associated with Impaired CD4 T-Cell Proliferation in Sepsis"

    Article Title: Upregulation of GRAIL Is Associated with Impaired CD4 T-Cell Proliferation in Sepsis

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1302160

    Upregulation of GRAIL in CD4 T-cells following sepsis (A) After 24 h of CLP, spleen tissues were harvested from shams and septic mice and fixed in 10% formalin and embedded in paraffin. Tissue blocks were sectioned at a thickness of 5 μm and incubated with rabbit anti-mouse GRAIL Ab, followed by incubation with biotinylated anti-rabbit IgG. Staining was developed by Vectastain ABC reagent and a diaminobenzidine kit. Representative immunostaing images at original magnification of ×100 and ×200 ( inset ) are shown. (B) Cells were isolated from spleens of sham and CLP animals; a total of 2 ×10 6 cells were used to extract mRNA and subsequent realtime PCR analysis using GRAIL primer. The expression of β-actin serves as an internal control. The data is expressed as fold induction in comparison to the shams. (C) Splenocytes (1 × 10 6 cells) from the shams and septic mice were stained for surface CD4 antigen and intracellular GRAIL using PerCP/Cy5.5-CD4 Ab, and rabbit anti-GRAIL primary Ab, respectively. A FITC-labeled anti-rabbit secondary Ab was used subsequently and then events were acquired in flow cytometer. (D) Bar diagram representing the mean fluoresce intensities of sham and septic samples are shown. Data are expressed as means ± SE (n=5 mice/group). * P
    Figure Legend Snippet: Upregulation of GRAIL in CD4 T-cells following sepsis (A) After 24 h of CLP, spleen tissues were harvested from shams and septic mice and fixed in 10% formalin and embedded in paraffin. Tissue blocks were sectioned at a thickness of 5 μm and incubated with rabbit anti-mouse GRAIL Ab, followed by incubation with biotinylated anti-rabbit IgG. Staining was developed by Vectastain ABC reagent and a diaminobenzidine kit. Representative immunostaing images at original magnification of ×100 and ×200 ( inset ) are shown. (B) Cells were isolated from spleens of sham and CLP animals; a total of 2 ×10 6 cells were used to extract mRNA and subsequent realtime PCR analysis using GRAIL primer. The expression of β-actin serves as an internal control. The data is expressed as fold induction in comparison to the shams. (C) Splenocytes (1 × 10 6 cells) from the shams and septic mice were stained for surface CD4 antigen and intracellular GRAIL using PerCP/Cy5.5-CD4 Ab, and rabbit anti-GRAIL primary Ab, respectively. A FITC-labeled anti-rabbit secondary Ab was used subsequently and then events were acquired in flow cytometer. (D) Bar diagram representing the mean fluoresce intensities of sham and septic samples are shown. Data are expressed as means ± SE (n=5 mice/group). * P

    Techniques Used: Mouse Assay, Incubation, Staining, Isolation, Polymerase Chain Reaction, Expressing, Labeling, Flow Cytometry, Cytometry

    Related Articles

    Immunohistochemistry:

    Article Title: MyoD-Cre driven alterations in K-Ras and p53 lead to a mouse model with histological and molecular characteristics of human rhabdomyosarcoma with direct translational applications
    Article Snippet: .. For IHC, VECTASTAIN ABC Kit, Rabbit IgG (Vector Laboratories, CA, USA) was used, and the stainings were visualized using Vector NovaRED Peroxidase Substrate Kit (Vector Laboratories). ..

    Plasmid Preparation:

    Article Title: MyoD-Cre driven alterations in K-Ras and p53 lead to a mouse model with histological and molecular characteristics of human rhabdomyosarcoma with direct translational applications
    Article Snippet: .. For IHC, VECTASTAIN ABC Kit, Rabbit IgG (Vector Laboratories, CA, USA) was used, and the stainings were visualized using Vector NovaRED Peroxidase Substrate Kit (Vector Laboratories). ..

    Article Title: The Protein-Independent Role of Phosphate in the Progression of Chronic Kidney Disease
    Article Snippet: .. After being washed, sections were incubated with biotinylated anti-mouse IgG (H+L), (Vector, Burlingame, CA, USA) for 45 min, followed by VECTASTAIN® ABC-AP Kit, Vector® Red (Vector, Burlingame, CA, USA) for 30 min. ..

    Incubation:

    Article Title: Aspergillus versicolor Inhalation Triggers Neuroimmune, Glial, and Neuropeptide Transcriptional Changes
    Article Snippet: .. Sections were then washed three times in PBS, incubated with biotinylated anti-rabbit antibody for 1H, washed three times in PBS, and incubated with Vectastain ABC Kit (Vector Laboratories, Burlingame, CA) reagents according to manufacturer’s instructions. ..

    Article Title: The Protein-Independent Role of Phosphate in the Progression of Chronic Kidney Disease
    Article Snippet: .. After being washed, sections were incubated with biotinylated anti-mouse IgG (H+L), (Vector, Burlingame, CA, USA) for 45 min, followed by VECTASTAIN® ABC-AP Kit, Vector® Red (Vector, Burlingame, CA, USA) for 30 min. ..

    Article Title: Circulating activin A during equine gestation and immunolocalization of its receptors system in utero-placental tissues and fetal gonads
    Article Snippet: .. After incubation with the primary antibody, slides were rinsed for 5 min in PBS and incubated with a biotinylated second antibody for 30 min prior to incubation with ABC reagent for 30 min using a Vectastain ABC detection kit (Vector Laboratories). ..

    Article Title: Chondroprotective Effects of a Histone Deacetylase Inhibitor, Panobinostat, on Pain Behavior and Cartilage Degradation in Anterior Cruciate Ligament Transection-Induced Experimental Osteoarthritic Rats
    Article Snippet: .. Thereafter, sections were incubated with an avidin–biotin complex using an ABC kit (Vectastain ABC kit; Vector Laboratories Inc., Burlingame, CA, USA), followed by incubation with 3,3′-diaminobenzidine tetrahydrochloride for 5 min. ..

    Staining:

    Article Title: α-Synuclein pathology in Parkinson disease activates homeostatic NRF2 anti-oxidant response
    Article Snippet: .. The following antibodies were employed to stain serial tissue sections, as indicated: antibody against phospho-alpha synuclein (p-aSyn, S129; 81A monoclonal; EMD Millipore, #MABN826; dilution 1:1000) [ ], and antibody against phospho-NRF2 (p-NRF2, S40; EP1809Y monoclonal; abcam # ab76026); dilution 1:400) using the alkaline phosphatise conjugated streptavidin–biotin ABC kit (Vector Labs, #AK-5000). ..

    Article Title: Kv4.3 channel downregulation mediates chronic post-lesional pacemaker acceleration in surviving dopamine substantia nigra neurons
    Article Snippet: .. DAB immunocytochemistry For DAB (3,3′-diaminobenzidine) staining procedures, a Vectastain ABC Staining Kit (Vector Laboratories) was used. ..

    Article Title: Reversal of motor-skill transfer impairment by trihexyphenidyl and reduction of dorsolateral striatal cholinergic interneurons in Dyt1 ΔGAG knock-in mice
    Article Snippet: .. Every sixth section was stained with goat anti-ChAT antibody (EMD Millipore, AB144P; 1:100 dilution) with Vectastain ABC kit for peroxidase goat IgG and 3,3′-diaminobenzidine (DAB) peroxidase substrate kit (Vector laboratories). ..

    Immunocytochemistry:

    Article Title: Kv4.3 channel downregulation mediates chronic post-lesional pacemaker acceleration in surviving dopamine substantia nigra neurons
    Article Snippet: .. DAB immunocytochemistry For DAB (3,3′-diaminobenzidine) staining procedures, a Vectastain ABC Staining Kit (Vector Laboratories) was used. ..

    Avidin-Biotin Assay:

    Article Title: Chondroprotective Effects of a Histone Deacetylase Inhibitor, Panobinostat, on Pain Behavior and Cartilage Degradation in Anterior Cruciate Ligament Transection-Induced Experimental Osteoarthritic Rats
    Article Snippet: .. Thereafter, sections were incubated with an avidin–biotin complex using an ABC kit (Vectastain ABC kit; Vector Laboratories Inc., Burlingame, CA, USA), followed by incubation with 3,3′-diaminobenzidine tetrahydrochloride for 5 min. ..

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    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, 2021-09
    98/100 stars
      Buy from Supplier

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    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