detection kits  (Vector Laboratories)


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    Name:
    Vector TrueVIEW Autofluorescence Quenching Kit
    Description:
    The Vector TrueVIEW Autofluorescence Quenching Kit provides a novel way to diminish unwanted autofluorescence from non lipofuscin sources and dramatically improve signal to noise ratio It removes unwanted autofluorescence in tissue sections due to aldehyde fixation red blood cells and structural elements such as collagen and elastin The quenching action of the kit reagents provides a clear unambiguous “true view localization of the target antigen
    Catalog Number:
    SP-8400
    Price:
    None
    Category:
    Protein chemifluorescent detection reagents or kits or substrates
    Size:
    15 ml
    Buy from Supplier


    Structured Review

    Vector Laboratories detection kits
    The Vector TrueVIEW Autofluorescence Quenching Kit provides a novel way to diminish unwanted autofluorescence from non lipofuscin sources and dramatically improve signal to noise ratio It removes unwanted autofluorescence in tissue sections due to aldehyde fixation red blood cells and structural elements such as collagen and elastin The quenching action of the kit reagents provides a clear unambiguous “true view localization of the target antigen
    https://www.bioz.com/result/detection kits/product/Vector Laboratories
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    detection kits - by Bioz Stars, 2021-06
    97/100 stars

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    Related Articles

    Incubation:

    Article Title: Chronic Viral Infection Promotes Efficient Germinal Center B Cell Responses
    Article Snippet: Immunostaining was performed on 3 μm-thick sections using antibodies against GFP (ICL lab), Alexa Fluor 647-directly labeled B220 (eBioscience) and Lectin PNA Alexa Fluor 488 Conjugate (ThermoFisher). .. Immunostained slides were incubated with the Vector® TrueVIEW Autofluorescence Quenching Kit to remove autofluorescence signal (Vector Laboratories). .. Stained sections were scanned using a Panoramic Digital Slide Scanner 250 FLASH II (3DHISTECH) at 200 x magnification.

    Plasmid Preparation:

    Article Title: Chronic Viral Infection Promotes Efficient Germinal Center B Cell Responses
    Article Snippet: Immunostaining was performed on 3 μm-thick sections using antibodies against GFP (ICL lab), Alexa Fluor 647-directly labeled B220 (eBioscience) and Lectin PNA Alexa Fluor 488 Conjugate (ThermoFisher). .. Immunostained slides were incubated with the Vector® TrueVIEW Autofluorescence Quenching Kit to remove autofluorescence signal (Vector Laboratories). .. Stained sections were scanned using a Panoramic Digital Slide Scanner 250 FLASH II (3DHISTECH) at 200 x magnification.

    Article Title: Histone deacetylases 1 and 2 silence cryptic transcription to promote mitochondrial function during cardiogenesis
    Article Snippet: Eosin Y and Harris modified hematoxylin were obtained from Fisher, while X-gal was purchased from 5-Prime. .. Vectashield permanent mounting medium, Vectashield fluorescent mounting medium, Vectashield Elite ABC reagent kit, Vector TrueVIEW Autofluorescence Quenching Kit, and DAB Peroxidase Substrate kit were purchased from Vector Labs. .. ProLong Glass Antifade Mounting Medium was purchased from Thermo Fisher Scientific.

    Article Title: Presence of Clock genes in equine full-term placenta
    Article Snippet: .. Sections were rinsed three times with 1× PBS, then quenched for 5 min at room temperature with hydrogen peroxide (H2 O2) and Vector TrueView Autofluorescence Quenching Kit. ..

    Article Title: Slc1a3-2A-CreERT2 mice reveal unique features of Bergmann glia and augment a growing collection of Cre drivers and effectors in the 129S4 genetic background
    Article Snippet: Then, sections were washed in PBS for 3 × 5 min. After the last wash step secondary antibodies diluted in blocking buffer were added for additional 30 min incubation, RT. .. A second wash with PBS for 3 × 5 min, RT, was conducted prior to the use of a second auto fluorescence quenching kit (Vector TrueVIEW autofluorescence quenching kit, Vector Laboratories, # SP-8400). .. Finally, sections were washed in PBS for 5 min, incubated with DAPI (conc.

    Immunofluorescence:

    Article Title: Heme activates platelets and exacerbates rhabdomyolysis-induced acute kidney injury via CLEC-2 and GPVI/FcRγ
    Article Snippet: We incubated specimens with 1:200 rabbit anti-CitH3 antibody (Ab5103, Abcam, Cambridge, MA) and 1:200 rat anti-F4/80 antibody (eBioscience, San Diego, CA) at 4°C overnight and then with 1:1000 Alexa Fluor 555–labeled anti-rabbit IgG antibody (Abcam) and 1:500 Alexa Fluor 488–labeled anti–rat IgG antibody (Life Technologies, Carlsbad, CA) for 90 minutes. .. After immunofluorescence staining, autofluorescence was suppressed by using TrueVIEW (Vector Laboratories, Burlingame, CA). .. Finally, nuclear staining was performed with 5 µg/mL 4′,6-diamidino-2-phenylindole (Sigma-Aldrich) for 5 minutes.

    Staining:

    Article Title: Heme activates platelets and exacerbates rhabdomyolysis-induced acute kidney injury via CLEC-2 and GPVI/FcRγ
    Article Snippet: We incubated specimens with 1:200 rabbit anti-CitH3 antibody (Ab5103, Abcam, Cambridge, MA) and 1:200 rat anti-F4/80 antibody (eBioscience, San Diego, CA) at 4°C overnight and then with 1:1000 Alexa Fluor 555–labeled anti-rabbit IgG antibody (Abcam) and 1:500 Alexa Fluor 488–labeled anti–rat IgG antibody (Life Technologies, Carlsbad, CA) for 90 minutes. .. After immunofluorescence staining, autofluorescence was suppressed by using TrueVIEW (Vector Laboratories, Burlingame, CA). .. Finally, nuclear staining was performed with 5 µg/mL 4′,6-diamidino-2-phenylindole (Sigma-Aldrich) for 5 minutes.

    Fluorescence:

    Article Title: Slc1a3-2A-CreERT2 mice reveal unique features of Bergmann glia and augment a growing collection of Cre drivers and effectors in the 129S4 genetic background
    Article Snippet: Then, sections were washed in PBS for 3 × 5 min. After the last wash step secondary antibodies diluted in blocking buffer were added for additional 30 min incubation, RT. .. A second wash with PBS for 3 × 5 min, RT, was conducted prior to the use of a second auto fluorescence quenching kit (Vector TrueVIEW autofluorescence quenching kit, Vector Laboratories, # SP-8400). .. Finally, sections were washed in PBS for 5 min, incubated with DAPI (conc.

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  • 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, 2021-06
    98/100 stars
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    95
    Vector Laboratories immpress polymer anti mouse igg peroxidase reagent
    Mast cell activation in PD brains. Mid brain sections from PD patients and non-PD control subjects were incubated with 0.1% toluidine blue to detect mast cells (N=3). PD brains show increased mast cells (arrows, blue/purple metachromatic color) as well as increased activation as compared with non-PD control brains (a, b). Degranulated mast cells show extracellular granules and reduced blue color as seen in the PD brain. Further, we immunostained human mast cell specific tryptase in these brain sections using tryptase monoclonal antibody and ready-to-use <t>ImmPRESS</t> polymer anti-mouse <t>IgG</t> peroxidase reagent and ImmPACT DAB peroxidase substrate. Development of brown color indicated a positive reaction for tryptase ( c, d, arrows). Tryptase-positive mast cells and their activation status were increased in PD brains as compared to non-PD control subjects’ brains (c, d). Negative control staining performed without using the primary antibody did not show positive reaction for tryptase (e, f) . Original magnification = 200×.
    Immpress Polymer Anti Mouse Igg Peroxidase Reagent, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immpress polymer anti mouse igg peroxidase reagent/product/Vector Laboratories
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    immpress polymer anti mouse igg peroxidase reagent - by Bioz Stars, 2021-06
    95/100 stars
      Buy from Supplier

    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

    Mast cell activation in PD brains. Mid brain sections from PD patients and non-PD control subjects were incubated with 0.1% toluidine blue to detect mast cells (N=3). PD brains show increased mast cells (arrows, blue/purple metachromatic color) as well as increased activation as compared with non-PD control brains (a, b). Degranulated mast cells show extracellular granules and reduced blue color as seen in the PD brain. Further, we immunostained human mast cell specific tryptase in these brain sections using tryptase monoclonal antibody and ready-to-use ImmPRESS polymer anti-mouse IgG peroxidase reagent and ImmPACT DAB peroxidase substrate. Development of brown color indicated a positive reaction for tryptase ( c, d, arrows). Tryptase-positive mast cells and their activation status were increased in PD brains as compared to non-PD control subjects’ brains (c, d). Negative control staining performed without using the primary antibody did not show positive reaction for tryptase (e, f) . Original magnification = 200×.

    Journal: Molecular neurobiology

    Article Title: Mast Cell Proteases Activate Astrocytes and Glia-Neurons and Release Interleukin-33 by Activating p38 and ERK1/2 MAPKs and NF-κB

    doi: 10.1007/s12035-018-1177-7

    Figure Lengend Snippet: Mast cell activation in PD brains. Mid brain sections from PD patients and non-PD control subjects were incubated with 0.1% toluidine blue to detect mast cells (N=3). PD brains show increased mast cells (arrows, blue/purple metachromatic color) as well as increased activation as compared with non-PD control brains (a, b). Degranulated mast cells show extracellular granules and reduced blue color as seen in the PD brain. Further, we immunostained human mast cell specific tryptase in these brain sections using tryptase monoclonal antibody and ready-to-use ImmPRESS polymer anti-mouse IgG peroxidase reagent and ImmPACT DAB peroxidase substrate. Development of brown color indicated a positive reaction for tryptase ( c, d, arrows). Tryptase-positive mast cells and their activation status were increased in PD brains as compared to non-PD control subjects’ brains (c, d). Negative control staining performed without using the primary antibody did not show positive reaction for tryptase (e, f) . Original magnification = 200×.

    Article Snippet: ImmPRESS polymer anti-mouse IgG peroxidase reagent (made in goat) kit, ImmPACT DAB peroxidase substrate kit and antifade mounting medium were purchased from Vector Laboratories (Burlingame, MA).

    Techniques: Activation Assay, Incubation, Negative Control, Staining