alkaline phosphatase Search Results


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  • 99
    Vector Laboratories vectastain abc kit
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Vectastain Abc Kit, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 26234 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs calf intestinal alkaline phosphatase
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Calf Intestinal Alkaline Phosphatase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2886 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore alkaline phosphatase
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Alkaline Phosphatase, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 8519 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher alkaline phosphatase
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Alkaline Phosphatase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 4016 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher shrimp alkaline phosphatase
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Shrimp Alkaline Phosphatase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 4320 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore alkaline phosphatase activity
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Alkaline Phosphatase Activity, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2773 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Promega alkaline phosphatase
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Alkaline Phosphatase, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 2578 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore alkaline phosphatase staining
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Alkaline Phosphatase Staining, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1265 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore alkaline phosphatase detection kit
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Alkaline Phosphatase Detection Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1357 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs shrimp alkaline phosphatase
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Shrimp Alkaline Phosphatase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1499 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher fastap thermosensitive alkaline phosphatase
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Fastap Thermosensitive Alkaline Phosphatase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1527 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Boehringer Mannheim alkaline phosphatase
    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a <t>Vectastain</t> <t>ABC</t> kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.
    Alkaline Phosphatase, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 93/100, based on 1231 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore alkaline phosphatase conjugated secondary antibody
    Transglutaminase binds to immobilized PLCδ1. Human erythrocyte TG (0.05–2 μg/well) was allowed to bind to wells coated with two layers of PLCδ1 (0.2 μg/well) or to BSA-coated wells. Bound TG was monitored by ELISA by using a mAb to TG (H68.1.1) and an <t>alkaline</t> <t>phosphatase-conjugated</t> <t>secondary</t> <t>antibody.</t>
    Alkaline Phosphatase Conjugated Secondary Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 714 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore anti rabbit igg whole molecule alkaline phosphatase antibody
    Immunogenicity of alga-produced <t>CtxB-Pfs25</t> and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific <t>IgG</t>
    Anti Rabbit Igg Whole Molecule Alkaline Phosphatase Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1080 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    GE Healthcare shrimp alkaline phosphatase
    Immunogenicity of alga-produced <t>CtxB-Pfs25</t> and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific <t>IgG</t>
    Shrimp Alkaline Phosphatase, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 94/100, based on 1683 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore alkaline phosphatase substrate
    Immunogenicity of alga-produced <t>CtxB-Pfs25</t> and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific <t>IgG</t>
    Alkaline Phosphatase Substrate, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1011 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore leukocyte alkaline phosphatase kit
    Immunogenicity of alga-produced <t>CtxB-Pfs25</t> and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific <t>IgG</t>
    Leukocyte Alkaline Phosphatase Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 932 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies alkaline phosphatase
    Immunogenicity of alga-produced <t>CtxB-Pfs25</t> and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific <t>IgG</t>
    Alkaline Phosphatase, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 1050 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad alkaline phosphatase
    Immunogenicity of alga-produced <t>CtxB-Pfs25</t> and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific <t>IgG</t>
    Alkaline Phosphatase, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1076 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa alkaline phosphatase
    Immunogenicity of alga-produced <t>CtxB-Pfs25</t> and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific <t>IgG</t>
    Alkaline Phosphatase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1045 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore anti mouse igg whole molecule alkaline phosphatase antibody
    MK-0524 induces translocation of DP1 from intracellular compartments to the plasma membrane. The distribution of DP1 in HEK293 cells was determined by immunofluorescence confocal microscopy. HEK293 cells transfected with <t>Flag-DP1</t> were treated with vehicle or 1 µM of MK-0524 alone or in presence of 20 µM Brefeldin A for 90 min. Cells were labeled with mouse anti-FLAG and either a rabbit anti-calnexin antibody (top and midlle panels) or a rabbit anti-protein disulfide isomerase (PDI) antibody (lower panel) as described under “ Materials and Methods ”. Secondary antibodies were Alexa Fluor 488 donkey anti-mouse <t>IgG</t> and Alexa Fluor 546 goat anti-rabbit IgG. Merge images of the green-labelled DP1 and red-labeled calnexin or PDI are shown. Bars, 10 µM.
    Anti Mouse Igg Whole Molecule Alkaline Phosphatase Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1128 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a Vectastain ABC kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.

    Journal: Reproduction (Cambridge, England)

    Article Title: Molecular characterization of bovine placental and ovarian 20?-hydroxysteroid dehydrogenase

    doi: 10.1530/REP-11-0093

    Figure Lengend Snippet: Localization of bovine 20α-HSD protein expression in the CL throughout the estrous cycle by immunohistochemistry. Representative immunohistochemical analyses using 20α-HSD antiserum (1:1000) and goat anti-rabbit secondary antibodies (1:500). Preimmune serum (1:1000) was used for primary antiserum as the negative control. Immunohistochemistry was performed by a Vectastain ABC kit. Ovarian sections are shown during the estrous cycle. Preimmune serum was used as the control for the CL stage 7 below the right panel. Black bar=100 μm. Red arrows indicate luteal cells.

    Article Snippet: Immunohistochemistry Immunohistochemical staining of CH2, CH3, CL3, CL2, and CL1 samples was performed by the Vectastain ABC kit (Vector Laboratories).

    Techniques: Expressing, Immunohistochemistry, Negative Control

    Localization of CD148 + CD27 + and CD148 − CD27 − B cells in follicles of human spleen. Serial tissue sections of human spleen were incubated with anti-IgD antiserum alone ( blue ; A and D ), anti-IgD antiserum ( blue ) and anti-CD27 mAb ( red ; B and E ), or anti-IgD antiserum ( blue ) and anti-IgM mAb ( red ; C ). The anti-IgD polyclonal antibody was visualized after the addition of alkaline phosphatase–conjugated anti–goat Ig and phosphatase-specific substrate. The anti-CD27 or anti-IgM mAb was visualized by an anti–mouse Ig-specific Vectastain kit. Original magnification of A , B , and C : ×40; of D and E : ×100. The follicular mantle zone ( FM ), marginal zone ( MZ ), germinal center ( G or GC ), and T cell zones ( T ) are indicated.

    Journal: The Journal of Experimental Medicine

    Article Title: Identification of Functional Human Splenic Memory B Cells by Expression of CD148 and CD27

    doi:

    Figure Lengend Snippet: Localization of CD148 + CD27 + and CD148 − CD27 − B cells in follicles of human spleen. Serial tissue sections of human spleen were incubated with anti-IgD antiserum alone ( blue ; A and D ), anti-IgD antiserum ( blue ) and anti-CD27 mAb ( red ; B and E ), or anti-IgD antiserum ( blue ) and anti-IgM mAb ( red ; C ). The anti-IgD polyclonal antibody was visualized after the addition of alkaline phosphatase–conjugated anti–goat Ig and phosphatase-specific substrate. The anti-CD27 or anti-IgM mAb was visualized by an anti–mouse Ig-specific Vectastain kit. Original magnification of A , B , and C : ×40; of D and E : ×100. The follicular mantle zone ( FM ), marginal zone ( MZ ), germinal center ( G or GC ), and T cell zones ( T ) are indicated.

    Article Snippet: Bound mouse mAb was visualized using a mouse Ig–specific Vectastain kit (Vector Laboratories, Inc., Burlingame, CA), according to the manufacturer's instructions.

    Techniques: Incubation

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    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: In some experiments, frozen sections were stained using a Vectastain ABC-AP kit (Vector Laboratories, AK-5000) containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase.

    Techniques: Isolation, Mouse Assay, Staining

    Transglutaminase binds to immobilized PLCδ1. Human erythrocyte TG (0.05–2 μg/well) was allowed to bind to wells coated with two layers of PLCδ1 (0.2 μg/well) or to BSA-coated wells. Bound TG was monitored by ELISA by using a mAb to TG (H68.1.1) and an alkaline phosphatase-conjugated secondary antibody.

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

    Article Title: Interactions of Gh/transglutaminase with phospholipase C?1 and with GTP

    doi:

    Figure Lengend Snippet: Transglutaminase binds to immobilized PLCδ1. Human erythrocyte TG (0.05–2 μg/well) was allowed to bind to wells coated with two layers of PLCδ1 (0.2 μg/well) or to BSA-coated wells. Bound TG was monitored by ELISA by using a mAb to TG (H68.1.1) and an alkaline phosphatase-conjugated secondary antibody.

    Article Snippet: After washing with 3 × 300 μl/well of 2% BSA, 125 μl/well alkaline phosphatase-conjugated secondary antibody at 1:5,000 dilution in 2% BSA (anti-mouse IgG developed in rabbit; Sigma) was added and allowed to bind for 2 hr.

    Techniques: Enzyme-linked Immunosorbent Assay

    Immunogenicity of alga-produced CtxB-Pfs25 and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific IgG

    Journal: Applied and Environmental Microbiology

    Article Title: Alga-Produced Cholera Toxin-Pfs25 Fusion Proteins as Oral Vaccines

    doi: 10.1128/AEM.00714-13

    Figure Lengend Snippet: Immunogenicity of alga-produced CtxB-Pfs25 and Pfs5 using Alhydrogel. Mice were vaccinated by intraperitoneal injection with 20 μg of affinity-purified Cr.CtxB-Pfs25 or the molar equivalent of Cr.Pfs25 adsorbed to Alhydrogel. Pfs25-specific IgG

    Article Snippet: The membranes were probed using rabbit anti-CT antibodies (Sigma catalog no. C3062), mouse anti-FLAG MAbs (Sigma catalog no. F3165), or anti-Pfs25 (4B7) MAbs and detected with goat anti-rabbit IgG (Sigma catalog no. A3687) or goat anti-mouse IgG (Sigma catalog no. A1682) secondary antibodies conjugated to alkaline phosphatase.

    Techniques: Produced, Mouse Assay, Injection, Affinity Purification

    Immune response to whole-cell alga oral immunization in naive mice. Mice were administered weekly doses of JAG101 algae or Δ psbA C. reinhardtii . Serum IgG antibodies for Pfs25 (A) and CtxB (B) were detected by ELISA. Error bars indicate standard

    Journal: Applied and Environmental Microbiology

    Article Title: Alga-Produced Cholera Toxin-Pfs25 Fusion Proteins as Oral Vaccines

    doi: 10.1128/AEM.00714-13

    Figure Lengend Snippet: Immune response to whole-cell alga oral immunization in naive mice. Mice were administered weekly doses of JAG101 algae or Δ psbA C. reinhardtii . Serum IgG antibodies for Pfs25 (A) and CtxB (B) were detected by ELISA. Error bars indicate standard

    Article Snippet: The membranes were probed using rabbit anti-CT antibodies (Sigma catalog no. C3062), mouse anti-FLAG MAbs (Sigma catalog no. F3165), or anti-Pfs25 (4B7) MAbs and detected with goat anti-rabbit IgG (Sigma catalog no. A3687) or goat anti-mouse IgG (Sigma catalog no. A1682) secondary antibodies conjugated to alkaline phosphatase.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    MK-0524 induces translocation of DP1 from intracellular compartments to the plasma membrane. The distribution of DP1 in HEK293 cells was determined by immunofluorescence confocal microscopy. HEK293 cells transfected with Flag-DP1 were treated with vehicle or 1 µM of MK-0524 alone or in presence of 20 µM Brefeldin A for 90 min. Cells were labeled with mouse anti-FLAG and either a rabbit anti-calnexin antibody (top and midlle panels) or a rabbit anti-protein disulfide isomerase (PDI) antibody (lower panel) as described under “ Materials and Methods ”. Secondary antibodies were Alexa Fluor 488 donkey anti-mouse IgG and Alexa Fluor 546 goat anti-rabbit IgG. Merge images of the green-labelled DP1 and red-labeled calnexin or PDI are shown. Bars, 10 µM.

    Journal: PLoS ONE

    Article Title: Inverse Agonist and Pharmacochaperone Properties of MK-0524 on the Prostanoid DP1 Receptor

    doi: 10.1371/journal.pone.0065767

    Figure Lengend Snippet: MK-0524 induces translocation of DP1 from intracellular compartments to the plasma membrane. The distribution of DP1 in HEK293 cells was determined by immunofluorescence confocal microscopy. HEK293 cells transfected with Flag-DP1 were treated with vehicle or 1 µM of MK-0524 alone or in presence of 20 µM Brefeldin A for 90 min. Cells were labeled with mouse anti-FLAG and either a rabbit anti-calnexin antibody (top and midlle panels) or a rabbit anti-protein disulfide isomerase (PDI) antibody (lower panel) as described under “ Materials and Methods ”. Secondary antibodies were Alexa Fluor 488 donkey anti-mouse IgG and Alexa Fluor 546 goat anti-rabbit IgG. Merge images of the green-labelled DP1 and red-labeled calnexin or PDI are shown. Bars, 10 µM.

    Article Snippet: Reagents Monoclonal anti-FLAG (M2) (cat. F3165), monoclonal anti-FLAG (M1) (cat. F3040), and goat alkaline phosphatase-conjugated anti-mouse IgG (cat. A3562) antibodies were from Sigma-Aldrich, MO.

    Techniques: Translocation Assay, Immunofluorescence, Confocal Microscopy, Transfection, Labeling