streptavidin labeled horseradish peroxidase  (Thermo Fisher)


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    Structured Review

    Thermo Fisher streptavidin labeled horseradish peroxidase
    Streptavidin Labeled Horseradish Peroxidase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin labeled horseradish peroxidase/product/Thermo Fisher
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    streptavidin labeled horseradish peroxidase - by Bioz Stars, 2020-03
    85/100 stars

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    Related Articles

    Amplification:

    Article Title: Exercise reduces GABA synaptic input onto NTS baroreceptor second-order neurons via NK1 receptor internalization in spontaneously hypertensive rats
    Article Snippet: Tyramide signal amplification (TSA) immunofluoresence was performed on free-floating tissue. .. First, tissue was incubated in biotinylated goat anti-rabbit secondary antibody (3 μg/ml, Vector Labs) for 100 min, followed by strepavidin labeled horseradish peroxidase (2.5 μg/ml SA-HRP, TSA kit #23, Molecular Probes) for 60 min. Staining was visualized with Alexa Fluor 546 labeled tyramide (1:100) diluted in amplfication diluent + 0.0015% hydrogen peroxide for 22 min (TSA kit, Molecular Probes).

    Sandwich ELISA:

    Article Title: N-Substituted Pyrrole Derivatives as Novel Human Immunodeficiency Virus Type 1 Entry Inhibitors That Interfere with the gp41 Six-Helix Bundle Formation and Block Virus Fusion
    Article Snippet: A sandwich ELISA as previously described ( ) was used to screen for compounds that inhibit the gp41 six-helix bundle formation. .. Then, MAb NC-1, biotin-labeled goat anti-mouse IgG (Sigma Chemical Co., St. Louis, Mo.), streptavidin-labeled horseradish peroxidase (SA-HRP; Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (TMB; Sigma) were added sequentially.

    Next-Generation Sequencing:

    Article Title: Exercise reduces GABA synaptic input onto NTS baroreceptor second-order neurons via NK1 receptor internalization in spontaneously hypertensive rats
    Article Snippet: Sections were blocked in 1% blocking reagent (TSA kit, Molecular probes) and 10% normal goat serum (NGS, Vector Labs) for 60 min, then incubated simultaneously in primary antibodies for the NK1-R (0.2 μg/ml polyclonal rabbit anti-NK1-R, Chemicon AB5897) and GAD-67 (0.5 μg/ml monoclonal mouse anti-GAD, Chemicon MAB5406) in PBS/1% block/1% NGS for 60-72 hr at 4°C. .. First, tissue was incubated in biotinylated goat anti-rabbit secondary antibody (3 μg/ml, Vector Labs) for 100 min, followed by strepavidin labeled horseradish peroxidase (2.5 μg/ml SA-HRP, TSA kit #23, Molecular Probes) for 60 min. Staining was visualized with Alexa Fluor 546 labeled tyramide (1:100) diluted in amplfication diluent + 0.0015% hydrogen peroxide for 22 min (TSA kit, Molecular Probes).

    Incubation:

    Article Title: Potent HIV fusion inhibitors against Enfuvirtide-resistant HIV-1 strains
    Article Snippet: The mixture was then added to the NC-1-coated plate, followed by incubation at room temperature for 30 min and washing with a washing buffer (PBS containing 0.1% Tween 20) three times. .. Then, streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: N-Substituted Pyrrole Derivatives as Novel Human Immunodeficiency Virus Type 1 Entry Inhibitors That Interfere with the gp41 Six-Helix Bundle Formation and Block Virus Fusion
    Article Snippet: After incubation at 37°C for 30 min, the mixture was added to wells of a 96-well polystyrene plate (Costar; Corning, Inc., Corning, N.Y.), which were precoated with IgG (2 μg/ml) purified from rabbit antisera directed against the N36-C34 mixture. .. Then, MAb NC-1, biotin-labeled goat anti-mouse IgG (Sigma Chemical Co., St. Louis, Mo.), streptavidin-labeled horseradish peroxidase (SA-HRP; Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (TMB; Sigma) were added sequentially.

    Article Title: Development and Preclinical Studies of Broad-spectrum Anti-HIV Agent (3?R,4?R)-3-Cyanomethyl-4-methyl-3?,4?-di-O-(S)-camphanoyl-(+)-cis-khellactone (3-Cyanomethyl-4-methyl-DCK)
    Article Snippet: .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3',5,5'-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially. ..

    Article Title: Discovery of Critical Residues for Viral Entry and Inhibition through Structural Insight of HIV-1 Fusion Inhibitor CP621-652 *
    Article Snippet: The mixture was then added to the NC-1-coated plate, followed by incubation at room temperature for 30 min and washing with a washing buffer (PBS containing 0.1% Tween 20) three times. .. Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Biophysical Property and Broad Anti-HIV Activity of Albuvirtide, a 3-Maleimimidopropionic Acid-Modified Peptide Fusion Inhibitor
    Article Snippet: The mixture was then added to the NC-1-coated plate, followed by incubation at room temperature for 30 min and washing with a washing buffer (PBS containing 0.1% Tween 20) three times. .. Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5- tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Novel Recombinant Engineered gp41 N-terminal Heptad Repeat Trimers and Their Potential as Anti-HIV-1 Therapeutics or Microbicides *
    Article Snippet: Peptide (40 μg/ml) was incubated at 37 °C in PBS containing 1 microunit/ml proteinase K-Acrylic Beads (Sigma). .. Then, biotin-labeled goat anti-rabbit IgG (Sigma), streptavidin-labeled horseradish peroxidase (SA-HRP, Zymed Laboratories Inc., South San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Identification of a Critical Motif for the Human Immunodeficiency Virus Type 1 (HIV-1) gp41 Core Structure: Implications for Designing Novel Anti-HIV Fusion Inhibitors
    Article Snippet: The mixture was then added to the NC-1-coated plate, followed by incubation at room temperature for 30 min and washing with a washing buffer (phosphate-buffered saline [PBS] containing 0.1% Tween 20) three times. .. Streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were then added sequentially.

    Article Title: HIV-1 Fusion Is Blocked through Binding of GB Virus C E2D Peptides to the HIV-1 gp41 Disulfide Loop
    Article Snippet: After incubation for 30 min at 37°C, C34-biotin (0.25 µM) was added, followed by incubation for 45 min at 37°C and washing with wash buffer (PBS containing 0.05% Tween-20) six times. .. Streptavidin-labeled horseradish peroxidase (Invitrogen, Grand Island, NY) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma, St. Louis, MO) were added sequentially.

    Article Title: Design, Synthesis, and Structure-Activity Relationship of a Novel Series of 2-Aryl 5-(4-oxo-3-phenethyl-2-thioxothiazolidin-ylidenemethyl)furans as HIV-1 entry inhibitors *
    Article Snippet: .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially. ..

    Article Title: Diarylaniline Derivatives as a Distinct Class of HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors
    Article Snippet: .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially. ..

    Article Title: Exercise reduces GABA synaptic input onto NTS baroreceptor second-order neurons via NK1 receptor internalization in spontaneously hypertensive rats
    Article Snippet: .. First, tissue was incubated in biotinylated goat anti-rabbit secondary antibody (3 μg/ml, Vector Labs) for 100 min, followed by strepavidin labeled horseradish peroxidase (2.5 μg/ml SA-HRP, TSA kit #23, Molecular Probes) for 60 min. Staining was visualized with Alexa Fluor 546 labeled tyramide (1:100) diluted in amplfication diluent + 0.0015% hydrogen peroxide for 22 min (TSA kit, Molecular Probes). .. Sections were incubated in 0.3% hydrogen peroxide for 30 min to quench remaining peroxidase activity of the SA-HRP, rinsed and incubated in biotinylated goat anti-mouse antibody for 100 min, followed by SA-HRP for 60 min.

    Labeling:

    Article Title: Development and Preclinical Studies of Broad-spectrum Anti-HIV Agent (3?R,4?R)-3-Cyanomethyl-4-methyl-3?,4?-di-O-(S)-camphanoyl-(+)-cis-khellactone (3-Cyanomethyl-4-methyl-DCK)
    Article Snippet: .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3',5,5'-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially. ..

    Article Title: Exercise reduces GABA synaptic input onto NTS baroreceptor second-order neurons via NK1 receptor internalization in spontaneously hypertensive rats
    Article Snippet: .. First, tissue was incubated in biotinylated goat anti-rabbit secondary antibody (3 μg/ml, Vector Labs) for 100 min, followed by strepavidin labeled horseradish peroxidase (2.5 μg/ml SA-HRP, TSA kit #23, Molecular Probes) for 60 min. Staining was visualized with Alexa Fluor 546 labeled tyramide (1:100) diluted in amplfication diluent + 0.0015% hydrogen peroxide for 22 min (TSA kit, Molecular Probes). .. Sections were incubated in 0.3% hydrogen peroxide for 30 min to quench remaining peroxidase activity of the SA-HRP, rinsed and incubated in biotinylated goat anti-mouse antibody for 100 min, followed by SA-HRP for 60 min.

    Mouse Assay:

    Article Title: A Conformation-Specific Monoclonal Antibody Reacting with Fusion-Active gp41 from the Human Immunodeficiency Virus Type 1 Envelope Glycoprotein
    Article Snippet: Immunoglobulin G (IgG) was purified from the ascites fluid obtained from mice injected with NC-1 hybridoma cells and was used for the immunological studies. .. Then, biotin-labeled goat anti-mouse IgG (Boehringer Mannheim, Indianapolis, Ind.), streptavidin-labeled horseradish peroxidase (Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, Mo.) were added sequentially.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Potent HIV fusion inhibitors against Enfuvirtide-resistant HIV-1 strains
    Article Snippet: Inhibitory activity of the peptides (CP32M, CP621–652, C34, and T20) on the 6-HB formation was measured by a modified ELISA-based method as previously described ( ). .. Then, streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: N-Substituted Pyrrole Derivatives as Novel Human Immunodeficiency Virus Type 1 Entry Inhibitors That Interfere with the gp41 Six-Helix Bundle Formation and Block Virus Fusion
    Article Snippet: Paragraph title: ELISA for screening for compounds that inhibit the gp41 six-helix bundle formation. ... Then, MAb NC-1, biotin-labeled goat anti-mouse IgG (Sigma Chemical Co., St. Louis, Mo.), streptavidin-labeled horseradish peroxidase (SA-HRP; Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (TMB; Sigma) were added sequentially.

    Article Title: Development and Preclinical Studies of Broad-spectrum Anti-HIV Agent (3?R,4?R)-3-Cyanomethyl-4-methyl-3?,4?-di-O-(S)-camphanoyl-(+)-cis-khellactone (3-Cyanomethyl-4-methyl-DCK)
    Article Snippet: On the fourth day post-infection, 100 µL of culture supernatants were collected from each well, mixed with equal volumes of 5% Triton X-100 and assayed for p24 antigen, which was quantitated by ELISA. .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3',5,5'-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Discovery of Critical Residues for Viral Entry and Inhibition through Structural Insight of HIV-1 Fusion Inhibitor CP621-652 *
    Article Snippet: The inhibitory activity of CP621–652 or its mutants (M626A and T627A) on 6-HB formation was measured by a modified ELISA-based method as previously described ( , ). .. Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Biophysical Property and Broad Anti-HIV Activity of Albuvirtide, a 3-Maleimimidopropionic Acid-Modified Peptide Fusion Inhibitor
    Article Snippet: Inhibitory activity of ABT, T20 and C34 on the 6-HB formation was measured by a modified ELISA-based method as previously described , . .. Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5- tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Identification of a Critical Motif for the Human Immunodeficiency Virus Type 1 (HIV-1) gp41 Core Structure: Implications for Designing Novel Anti-HIV Fusion Inhibitors
    Article Snippet: The inhibitory activities of peptides (CP621-652, C34, and T-20) on 6-HB formation were measured by a modified enzyme-linked immunosorbent assay (ELISA)-based method as previously described ( ). .. Streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were then added sequentially.

    Article Title: HIV-1 Fusion Is Blocked through Binding of GB Virus C E2D Peptides to the HIV-1 gp41 Disulfide Loop
    Article Snippet: Inhibition of 6-HB Formation Inhibitory activity of the peptides (P4-7, P6-2, and P28) on 6-HB formation was measured by a modified ELISA-based method as previously described . .. Streptavidin-labeled horseradish peroxidase (Invitrogen, Grand Island, NY) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma, St. Louis, MO) were added sequentially.

    Article Title: Design, Synthesis, and Structure-Activity Relationship of a Novel Series of 2-Aryl 5-(4-oxo-3-phenethyl-2-thioxothiazolidin-ylidenemethyl)furans as HIV-1 entry inhibitors *
    Article Snippet: On the fourth day post-infection, 100 μL of culture supernatants were collected from each well, mixed with equal volumes of 5% Triton X-100 and assayed for p24 antigen, which was quantitated by ELISA. .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Diarylaniline Derivatives as a Distinct Class of HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors
    Article Snippet: On the fourth day post-infection, 100 μL of culture supernatants were collected from each well, mixed with equal volumes of 5% Triton X-100 and assayed for p24 antigen, which was quantitated by ELISA. .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Concentration Assay:

    Article Title: N-Substituted Pyrrole Derivatives as Novel Human Immunodeficiency Virus Type 1 Entry Inhibitors That Interfere with the gp41 Six-Helix Bundle Formation and Block Virus Fusion
    Article Snippet: Then, MAb NC-1, biotin-labeled goat anti-mouse IgG (Sigma Chemical Co., St. Louis, Mo.), streptavidin-labeled horseradish peroxidase (SA-HRP; Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (TMB; Sigma) were added sequentially. .. The percent inhibition by the compounds was calculated as previously described ( ) and the effective concentration for 50% inhibition (EC50 ) was calculated by using the software Calcusyn , kindly provided by T. C. Chou (Sloan-Kettering Cancer Center, New York, N.Y.).

    Purification:

    Article Title: N-Substituted Pyrrole Derivatives as Novel Human Immunodeficiency Virus Type 1 Entry Inhibitors That Interfere with the gp41 Six-Helix Bundle Formation and Block Virus Fusion
    Article Snippet: After incubation at 37°C for 30 min, the mixture was added to wells of a 96-well polystyrene plate (Costar; Corning, Inc., Corning, N.Y.), which were precoated with IgG (2 μg/ml) purified from rabbit antisera directed against the N36-C34 mixture. .. Then, MAb NC-1, biotin-labeled goat anti-mouse IgG (Sigma Chemical Co., St. Louis, Mo.), streptavidin-labeled horseradish peroxidase (SA-HRP; Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (TMB; Sigma) were added sequentially.

    Article Title: A Conformation-Specific Monoclonal Antibody Reacting with Fusion-Active gp41 from the Human Immunodeficiency Virus Type 1 Envelope Glycoprotein
    Article Snippet: Mouse sera and culture supernatants containing antibodies or purified IgG were added to the wells at various concentrations. .. Then, biotin-labeled goat anti-mouse IgG (Boehringer Mannheim, Indianapolis, Ind.), streptavidin-labeled horseradish peroxidase (Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, Mo.) were added sequentially.

    Inhibition:

    Article Title: Potent HIV fusion inhibitors against Enfuvirtide-resistant HIV-1 strains
    Article Snippet: Paragraph title: Inhibition of CP32M on 6-HB Formation. ... Then, streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: N-Substituted Pyrrole Derivatives as Novel Human Immunodeficiency Virus Type 1 Entry Inhibitors That Interfere with the gp41 Six-Helix Bundle Formation and Block Virus Fusion
    Article Snippet: Then, MAb NC-1, biotin-labeled goat anti-mouse IgG (Sigma Chemical Co., St. Louis, Mo.), streptavidin-labeled horseradish peroxidase (SA-HRP; Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (TMB; Sigma) were added sequentially. .. The percent inhibition by the compounds was calculated as previously described ( ) and the effective concentration for 50% inhibition (EC50 ) was calculated by using the software Calcusyn , kindly provided by T. C. Chou (Sloan-Kettering Cancer Center, New York, N.Y.).

    Article Title: Discovery of Critical Residues for Viral Entry and Inhibition through Structural Insight of HIV-1 Fusion Inhibitor CP621-652 *
    Article Snippet: Paragraph title: Inhibition of 6-HB Formation by Peptides ... Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Biophysical Property and Broad Anti-HIV Activity of Albuvirtide, a 3-Maleimimidopropionic Acid-Modified Peptide Fusion Inhibitor
    Article Snippet: Paragraph title: Inhibition of 6-HB formation by peptides ... Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5- tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Identification of a Critical Motif for the Human Immunodeficiency Virus Type 1 (HIV-1) gp41 Core Structure: Implications for Designing Novel Anti-HIV Fusion Inhibitors
    Article Snippet: Paragraph title: Inhibition of HIV-1 gp41 6-HB formation by CP621-652. ... Streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were then added sequentially.

    Article Title: HIV-1 Fusion Is Blocked through Binding of GB Virus C E2D Peptides to the HIV-1 gp41 Disulfide Loop
    Article Snippet: Paragraph title: Inhibition of 6-HB Formation ... Streptavidin-labeled horseradish peroxidase (Invitrogen, Grand Island, NY) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma, St. Louis, MO) were added sequentially.

    Activity Assay:

    Article Title: Potent HIV fusion inhibitors against Enfuvirtide-resistant HIV-1 strains
    Article Snippet: Inhibitory activity of the peptides (CP32M, CP621–652, C34, and T20) on the 6-HB formation was measured by a modified ELISA-based method as previously described ( ). .. Then, streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Development and Preclinical Studies of Broad-spectrum Anti-HIV Agent (3?R,4?R)-3-Cyanomethyl-4-methyl-3?,4?-di-O-(S)-camphanoyl-(+)-cis-khellactone (3-Cyanomethyl-4-methyl-DCK)
    Article Snippet: The inhibitory activity of compounds on HIV-1 infection was determined as previously described. .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3',5,5'-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Discovery of Critical Residues for Viral Entry and Inhibition through Structural Insight of HIV-1 Fusion Inhibitor CP621-652 *
    Article Snippet: The inhibitory activity of CP621–652 or its mutants (M626A and T627A) on 6-HB formation was measured by a modified ELISA-based method as previously described ( , ). .. Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Biophysical Property and Broad Anti-HIV Activity of Albuvirtide, a 3-Maleimimidopropionic Acid-Modified Peptide Fusion Inhibitor
    Article Snippet: Inhibitory activity of ABT, T20 and C34 on the 6-HB formation was measured by a modified ELISA-based method as previously described , . .. Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5- tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Novel Recombinant Engineered gp41 N-terminal Heptad Repeat Trimers and Their Potential as Anti-HIV-1 Therapeutics or Microbicides *
    Article Snippet: Then, biotin-labeled goat anti-rabbit IgG (Sigma), streptavidin-labeled horseradish peroxidase (SA-HRP, Zymed Laboratories Inc., South San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially. .. The remaining antiviral activity in the supernatants against HIV-1IIIB infection was determined as described above.

    Article Title: HIV-1 Fusion Is Blocked through Binding of GB Virus C E2D Peptides to the HIV-1 gp41 Disulfide Loop
    Article Snippet: Inhibition of 6-HB Formation Inhibitory activity of the peptides (P4-7, P6-2, and P28) on 6-HB formation was measured by a modified ELISA-based method as previously described . .. Streptavidin-labeled horseradish peroxidase (Invitrogen, Grand Island, NY) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma, St. Louis, MO) were added sequentially.

    Article Title: Design, Synthesis, and Structure-Activity Relationship of a Novel Series of 2-Aryl 5-(4-oxo-3-phenethyl-2-thioxothiazolidin-ylidenemethyl)furans as HIV-1 entry inhibitors *
    Article Snippet: Paragraph title: Determination of the inhibitory activity of the compounds on HIV-1 replication ... Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Diarylaniline Derivatives as a Distinct Class of HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors
    Article Snippet: Paragraph title: Assay for Measuring the Inhibitory Activity of Compounds on HIV-1 IIIB Replication in MT-2 Cells ... Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Exercise reduces GABA synaptic input onto NTS baroreceptor second-order neurons via NK1 receptor internalization in spontaneously hypertensive rats
    Article Snippet: First, tissue was incubated in biotinylated goat anti-rabbit secondary antibody (3 μg/ml, Vector Labs) for 100 min, followed by strepavidin labeled horseradish peroxidase (2.5 μg/ml SA-HRP, TSA kit #23, Molecular Probes) for 60 min. Staining was visualized with Alexa Fluor 546 labeled tyramide (1:100) diluted in amplfication diluent + 0.0015% hydrogen peroxide for 22 min (TSA kit, Molecular Probes). .. Sections were incubated in 0.3% hydrogen peroxide for 30 min to quench remaining peroxidase activity of the SA-HRP, rinsed and incubated in biotinylated goat anti-mouse antibody for 100 min, followed by SA-HRP for 60 min.

    Infection:

    Article Title: Development and Preclinical Studies of Broad-spectrum Anti-HIV Agent (3?R,4?R)-3-Cyanomethyl-4-methyl-3?,4?-di-O-(S)-camphanoyl-(+)-cis-khellactone (3-Cyanomethyl-4-methyl-DCK)
    Article Snippet: In brief, 1 × 104 MT-2 cells were infected with an HIV-1 strain (100 TCID50 ) in 200 µL RPMI 1640 medium containing 10% FBS in the presence or absence of a test compound at graded concentrations overnight. .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3',5,5'-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Novel Recombinant Engineered gp41 N-terminal Heptad Repeat Trimers and Their Potential as Anti-HIV-1 Therapeutics or Microbicides *
    Article Snippet: Then, biotin-labeled goat anti-rabbit IgG (Sigma), streptavidin-labeled horseradish peroxidase (SA-HRP, Zymed Laboratories Inc., South San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially. .. The remaining antiviral activity in the supernatants against HIV-1IIIB infection was determined as described above.

    Article Title: Design, Synthesis, and Structure-Activity Relationship of a Novel Series of 2-Aryl 5-(4-oxo-3-phenethyl-2-thioxothiazolidin-ylidenemethyl)furans as HIV-1 entry inhibitors *
    Article Snippet: In brief, 1×104 MT-2 cells were infected with an HIV-1IIIB strain (100 TCID50 ) in 200 μL RPMI 1640 medium containing 10% FBS in the presence or absence of a test compound at graded concentrations overnight. .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Diarylaniline Derivatives as a Distinct Class of HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors
    Article Snippet: In brief, 1 × 104 MT-2 cells were infected with an HIV-1 strain (100 TCID50 ) in 200 μL RPMI 1640 medium containing 10% FBS in the presence or absence of a test compound at graded concentrations overnight. .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Recombinant:

    Article Title: Development and Preclinical Studies of Broad-spectrum Anti-HIV Agent (3?R,4?R)-3-Cyanomethyl-4-methyl-3?,4?-di-O-(S)-camphanoyl-(+)-cis-khellactone (3-Cyanomethyl-4-methyl-DCK)
    Article Snippet: Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3',5,5'-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially. .. Recombinant protein p24 purchased from US Biological (Swampscott, MA) was included for establishing standard dose response curves.

    Article Title: Design, Synthesis, and Structure-Activity Relationship of a Novel Series of 2-Aryl 5-(4-oxo-3-phenethyl-2-thioxothiazolidin-ylidenemethyl)furans as HIV-1 entry inhibitors *
    Article Snippet: Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially. .. Recombinant protein p24 purchased from US Biological (Swampscott, MA) was included for establishing standard dose response curves.

    Article Title: Diarylaniline Derivatives as a Distinct Class of HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors
    Article Snippet: Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially. .. Recombinant protein p24 purchased from US Biological (Swampscott, MA) was included for establishing standard dose response curves.

    Modification:

    Article Title: Potent HIV fusion inhibitors against Enfuvirtide-resistant HIV-1 strains
    Article Snippet: Inhibitory activity of the peptides (CP32M, CP621–652, C34, and T20) on the 6-HB formation was measured by a modified ELISA-based method as previously described ( ). .. Then, streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Discovery of Critical Residues for Viral Entry and Inhibition through Structural Insight of HIV-1 Fusion Inhibitor CP621-652 *
    Article Snippet: The inhibitory activity of CP621–652 or its mutants (M626A and T627A) on 6-HB formation was measured by a modified ELISA-based method as previously described ( , ). .. Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5-tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Biophysical Property and Broad Anti-HIV Activity of Albuvirtide, a 3-Maleimimidopropionic Acid-Modified Peptide Fusion Inhibitor
    Article Snippet: Inhibitory activity of ABT, T20 and C34 on the 6-HB formation was measured by a modified ELISA-based method as previously described , . .. Then streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3,5,5- tetramethylbenzidine (Sigma) were added sequentially.

    Article Title: Identification of a Critical Motif for the Human Immunodeficiency Virus Type 1 (HIV-1) gp41 Core Structure: Implications for Designing Novel Anti-HIV Fusion Inhibitors
    Article Snippet: The inhibitory activities of peptides (CP621-652, C34, and T-20) on 6-HB formation were measured by a modified enzyme-linked immunosorbent assay (ELISA)-based method as previously described ( ). .. Streptavidin-labeled horseradish peroxidase (Invitrogen) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were then added sequentially.

    Article Title: HIV-1 Fusion Is Blocked through Binding of GB Virus C E2D Peptides to the HIV-1 gp41 Disulfide Loop
    Article Snippet: Inhibition of 6-HB Formation Inhibitory activity of the peptides (P4-7, P6-2, and P28) on 6-HB formation was measured by a modified ELISA-based method as previously described . .. Streptavidin-labeled horseradish peroxidase (Invitrogen, Grand Island, NY) and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma, St. Louis, MO) were added sequentially.

    Staining:

    Article Title: Exercise reduces GABA synaptic input onto NTS baroreceptor second-order neurons via NK1 receptor internalization in spontaneously hypertensive rats
    Article Snippet: .. First, tissue was incubated in biotinylated goat anti-rabbit secondary antibody (3 μg/ml, Vector Labs) for 100 min, followed by strepavidin labeled horseradish peroxidase (2.5 μg/ml SA-HRP, TSA kit #23, Molecular Probes) for 60 min. Staining was visualized with Alexa Fluor 546 labeled tyramide (1:100) diluted in amplfication diluent + 0.0015% hydrogen peroxide for 22 min (TSA kit, Molecular Probes). .. Sections were incubated in 0.3% hydrogen peroxide for 30 min to quench remaining peroxidase activity of the SA-HRP, rinsed and incubated in biotinylated goat anti-mouse antibody for 100 min, followed by SA-HRP for 60 min.

    Injection:

    Article Title: A Conformation-Specific Monoclonal Antibody Reacting with Fusion-Active gp41 from the Human Immunodeficiency Virus Type 1 Envelope Glycoprotein
    Article Snippet: Immunoglobulin G (IgG) was purified from the ascites fluid obtained from mice injected with NC-1 hybridoma cells and was used for the immunological studies. .. Then, biotin-labeled goat anti-mouse IgG (Boehringer Mannheim, Indianapolis, Ind.), streptavidin-labeled horseradish peroxidase (Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, Mo.) were added sequentially.

    Binding Assay:

    Article Title: Novel Recombinant Engineered gp41 N-terminal Heptad Repeat Trimers and Their Potential as Anti-HIV-1 Therapeutics or Microbicides *
    Article Snippet: In brief, 50 μl of sample was coated onto wells of a 96-well polystyrene plate, followed by addition of rabbit antibodies directed against the HIV-1 gp41 N- and C-peptide mixture ( ) for binding N28Fd (at 1:400 dilution) and T20 (at 1:2000 dilution), respectively. .. Then, biotin-labeled goat anti-rabbit IgG (Sigma), streptavidin-labeled horseradish peroxidase (SA-HRP, Zymed Laboratories Inc., South San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma) were added sequentially.

    Blocking Assay:

    Article Title: Development and Preclinical Studies of Broad-spectrum Anti-HIV Agent (3?R,4?R)-3-Cyanomethyl-4-methyl-3?,4?-di-O-(S)-camphanoyl-(+)-cis-khellactone (3-Cyanomethyl-4-methyl-DCK)
    Article Snippet: Briefly, wells of polystyrene plates (Immulon 1B, Dynex Technology, Chantilly, VA) were coated with HIV immunoglobulin (HIVIG), which was prepared from plasma of HIV-seropositive donors with high neutralizing titers against HIV-1IIIB , in 0.085 M carbonate-bicarbonate buffer (pH 9.6) at 4 °C overnight, followed by washes with washing buffer (0.01M PBS containing 0.05% Tween-20) and blocking with PBS containing 1% dry fat-free milk (Bio-Rad Inc., Hercules, CA). .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3',5,5'-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: A Conformation-Specific Monoclonal Antibody Reacting with Fusion-Active gp41 from the Human Immunodeficiency Virus Type 1 Envelope Glycoprotein
    Article Snippet: Briefly, a peptide or protein antigen dissolved in 0.1 M Tris (pH 8.8) was used to coat wells of a 96-well polystyrene plate (Immulon II; Dynatech Laboratories, Inc., Chantilly, Va.) and post-coat blocked with a blocking buffer (phosphate-buffered saline plus 5% horse serum). .. Then, biotin-labeled goat anti-mouse IgG (Boehringer Mannheim, Indianapolis, Ind.), streptavidin-labeled horseradish peroxidase (Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, Mo.) were added sequentially.

    Article Title: Design, Synthesis, and Structure-Activity Relationship of a Novel Series of 2-Aryl 5-(4-oxo-3-phenethyl-2-thioxothiazolidin-ylidenemethyl)furans as HIV-1 entry inhibitors *
    Article Snippet: Briefly, wells of polystyrene plates (Immulon 1B, Dynex Technology, Chantilly, VA) were coated with HIV immunoglobulin (HIVIG), which was prepared from plasma of HIV-seropositive donors with high neutralizing titers against HIV-1IIIB , in 0.085 M carbonate-bicarbonate buffer (pH 9.6) at 4 °C overnight, followed by washing with buffer (0.01 M PBS containing 0.05% Tween-20) and blocking with PBS containing 1% dry fat-free milk (Bio-Rad, Inc., Hercules, CA). .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Diarylaniline Derivatives as a Distinct Class of HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors
    Article Snippet: Briefly, wells of polystyrene plates (Immulon 1B, Dynex Technology, Chantilly, VA) were coated with HIV immunoglobulin (HIVIG) in 0.085 M carbonate-bicarbonate buffer (pH 9.6) at 4 °C overnight, followed by washes with washing buffer (0.01M PBS containing 0.05% Tween-20) and blocking with PBS containing 1% dry fat-free milk (Bio-Rad, Inc., Hercules, CA). .. Virus lysates were added to the wells and incubated at 37 °C for 1 h. After extensive washes, anti-p24 mAb (183-12H-5C), biotin-labeled anti-mouse IgG1 (Santa Cruz Biotech., Santa Cruz, CA), streptavidin-labeled horseradish peroxidase (Zymed, S. San Francisco, CA), and the substrate 3,3′,5,5′-tetramethylbenzidine (Sigma Chemical Co., St. Louis, MO) were added sequentially.

    Article Title: Exercise reduces GABA synaptic input onto NTS baroreceptor second-order neurons via NK1 receptor internalization in spontaneously hypertensive rats
    Article Snippet: Sections were blocked in 1% blocking reagent (TSA kit, Molecular probes) and 10% normal goat serum (NGS, Vector Labs) for 60 min, then incubated simultaneously in primary antibodies for the NK1-R (0.2 μg/ml polyclonal rabbit anti-NK1-R, Chemicon AB5897) and GAD-67 (0.5 μg/ml monoclonal mouse anti-GAD, Chemicon MAB5406) in PBS/1% block/1% NGS for 60-72 hr at 4°C. .. First, tissue was incubated in biotinylated goat anti-rabbit secondary antibody (3 μg/ml, Vector Labs) for 100 min, followed by strepavidin labeled horseradish peroxidase (2.5 μg/ml SA-HRP, TSA kit #23, Molecular Probes) for 60 min. Staining was visualized with Alexa Fluor 546 labeled tyramide (1:100) diluted in amplfication diluent + 0.0015% hydrogen peroxide for 22 min (TSA kit, Molecular Probes).

    Software:

    Article Title: N-Substituted Pyrrole Derivatives as Novel Human Immunodeficiency Virus Type 1 Entry Inhibitors That Interfere with the gp41 Six-Helix Bundle Formation and Block Virus Fusion
    Article Snippet: Then, MAb NC-1, biotin-labeled goat anti-mouse IgG (Sigma Chemical Co., St. Louis, Mo.), streptavidin-labeled horseradish peroxidase (SA-HRP; Zymed, South San Francisco, Calif.), and the substrate 3,3′,5,5′-tetramethylbenzidine (TMB; Sigma) were added sequentially. .. The percent inhibition by the compounds was calculated as previously described ( ) and the effective concentration for 50% inhibition (EC50 ) was calculated by using the software Calcusyn , kindly provided by T. C. Chou (Sloan-Kettering Cancer Center, New York, N.Y.).

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    Thermo Fisher peroxidase labeled streptavidin
    Peroxidase Labeled Streptavidin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher hrp labeled streptavidin
    Cry4Ba binding to the larval midgut epithelium and toxin proteolytic processing in the larvae. (A) Larval tissue sections were covered by Cry4Ba wild type (Cry4Ba) and mutants (indicated). A bound toxin was detected with antibodies C4bPAbs, this was followed by secondary antibodies; biotin conjugated anti-mouse IgG <t>/streptavidin-HRP</t> (see Methods). The dark-brown color represents toxin binding at BBM (arrow heads) from 3-independent experiments. A control is a larval section which was reacted with antibodies, but without toxin reaction (control). (B) Western blot analysis of a toxin associated with PM or gut in the larvae treated with Cry4Ba wild type (lane 2) and R158A (lane 3) with Cry4Ba specific antibodies C4bPAbs. Antibodies show negative reactivity with proteins which were isolated from the PMs and guts of normal larvae (lane 1). (C) Reactivity of the antibodies C4bPAbs with Y170A protein associated with gut (lane 1) and PM (lane 2). The 47 kDa and 18-20 kDa of the trypsin activated Cry4Ba in vitro (B-lane 4, C-lane 3) were used as positive controls.
    Hrp Labeled Streptavidin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp labeled streptavidin/product/Thermo Fisher
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    95
    Thermo Fisher horseradish peroxidase hrp labeled streptavidin
    Cry4Ba binding to the larval midgut epithelium and toxin proteolytic processing in the larvae. (A) Larval tissue sections were covered by Cry4Ba wild type (Cry4Ba) and mutants (indicated). A bound toxin was detected with antibodies C4bPAbs, this was followed by secondary antibodies; biotin conjugated anti-mouse IgG <t>/streptavidin-HRP</t> (see Methods). The dark-brown color represents toxin binding at BBM (arrow heads) from 3-independent experiments. A control is a larval section which was reacted with antibodies, but without toxin reaction (control). (B) Western blot analysis of a toxin associated with PM or gut in the larvae treated with Cry4Ba wild type (lane 2) and R158A (lane 3) with Cry4Ba specific antibodies C4bPAbs. Antibodies show negative reactivity with proteins which were isolated from the PMs and guts of normal larvae (lane 1). (C) Reactivity of the antibodies C4bPAbs with Y170A protein associated with gut (lane 1) and PM (lane 2). The 47 kDa and 18-20 kDa of the trypsin activated Cry4Ba in vitro (B-lane 4, C-lane 3) were used as positive controls.
    Horseradish Peroxidase Hrp Labeled Streptavidin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Cry4Ba binding to the larval midgut epithelium and toxin proteolytic processing in the larvae. (A) Larval tissue sections were covered by Cry4Ba wild type (Cry4Ba) and mutants (indicated). A bound toxin was detected with antibodies C4bPAbs, this was followed by secondary antibodies; biotin conjugated anti-mouse IgG /streptavidin-HRP (see Methods). The dark-brown color represents toxin binding at BBM (arrow heads) from 3-independent experiments. A control is a larval section which was reacted with antibodies, but without toxin reaction (control). (B) Western blot analysis of a toxin associated with PM or gut in the larvae treated with Cry4Ba wild type (lane 2) and R158A (lane 3) with Cry4Ba specific antibodies C4bPAbs. Antibodies show negative reactivity with proteins which were isolated from the PMs and guts of normal larvae (lane 1). (C) Reactivity of the antibodies C4bPAbs with Y170A protein associated with gut (lane 1) and PM (lane 2). The 47 kDa and 18-20 kDa of the trypsin activated Cry4Ba in vitro (B-lane 4, C-lane 3) were used as positive controls.

    Journal: BMB Reports

    Article Title: Functional characterizations of residues Arg-158 and Tyr-170 of the mosquito-larvicidal Bacillus thuringiensis Cry4Ba

    doi: 10.5483/BMBRep.2014.47.10.192

    Figure Lengend Snippet: Cry4Ba binding to the larval midgut epithelium and toxin proteolytic processing in the larvae. (A) Larval tissue sections were covered by Cry4Ba wild type (Cry4Ba) and mutants (indicated). A bound toxin was detected with antibodies C4bPAbs, this was followed by secondary antibodies; biotin conjugated anti-mouse IgG /streptavidin-HRP (see Methods). The dark-brown color represents toxin binding at BBM (arrow heads) from 3-independent experiments. A control is a larval section which was reacted with antibodies, but without toxin reaction (control). (B) Western blot analysis of a toxin associated with PM or gut in the larvae treated with Cry4Ba wild type (lane 2) and R158A (lane 3) with Cry4Ba specific antibodies C4bPAbs. Antibodies show negative reactivity with proteins which were isolated from the PMs and guts of normal larvae (lane 1). (C) Reactivity of the antibodies C4bPAbs with Y170A protein associated with gut (lane 1) and PM (lane 2). The 47 kDa and 18-20 kDa of the trypsin activated Cry4Ba in vitro (B-lane 4, C-lane 3) were used as positive controls.

    Article Snippet: Dark blue-dots were developed by an hour-long reaction with 1:8,000 of biotin conjugated rabbit anti-mouse IgG (Thermo Scientific), 1:50,000 dilution of HRP labeled streptavidin (Thermo Scientific) and reaction with 3'-diaminobenzidine; DAB substrate kit (Vector Laboratories, Burlingame, CA).

    Techniques: Binding Assay, Western Blot, Isolation, In Vitro