primary streptavidin horseradish peroxidase  (Thermo Fisher)


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    Structured Review

    Thermo Fisher primary streptavidin horseradish peroxidase
    Primary Streptavidin Horseradish Peroxidase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary streptavidin horseradish peroxidase/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary streptavidin horseradish peroxidase - by Bioz Stars, 2020-04
    86/100 stars

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    Related Articles

    Activity Assay:

    Article Title: The VASCULATURE COMPLEXITY AND CONNECTIVITY Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 [C] Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 [C] [W] Gene Encodes a Plant-Specific Protein Required for Embryo Provasculatur
    Article Snippet: Sections were then washed once in 0.1× SSC (3 m NaCl and 0.3 m sodium citrate, pH 7) and twice stringently with 0.1× SSC for 15 min at 42°C before treatment with 0.3% (v/v) in methanol for 30 min to block endogenous peroxidase activity. .. For signal detection, a 1:500-fold diluted primary streptavidin-horseradish peroxidase (0.5 mg mL−1 ; Thermo Scientific) solution was applied for 30 min on each section at room temperature.

    In Situ Hybridization:

    Article Title: The VASCULATURE COMPLEXITY AND CONNECTIVITY Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 [C] Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 [C] [W] Gene Encodes a Plant-Specific Protein Required for Embryo Provasculatur
    Article Snippet: Paragraph title: In Situ Hybridization ... For signal detection, a 1:500-fold diluted primary streptavidin-horseradish peroxidase (0.5 mg mL−1 ; Thermo Scientific) solution was applied for 30 min on each section at room temperature.

    Blocking Assay:

    Article Title: The VASCULATURE COMPLEXITY AND CONNECTIVITY Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 [C] Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 [C] [W] Gene Encodes a Plant-Specific Protein Required for Embryo Provasculatur
    Article Snippet: Sections were then washed once in 0.1× SSC (3 m NaCl and 0.3 m sodium citrate, pH 7) and twice stringently with 0.1× SSC for 15 min at 42°C before treatment with 0.3% (v/v) in methanol for 30 min to block endogenous peroxidase activity. .. For signal detection, a 1:500-fold diluted primary streptavidin-horseradish peroxidase (0.5 mg mL−1 ; Thermo Scientific) solution was applied for 30 min on each section at room temperature.

    Hybridization:

    Article Title: The VASCULATURE COMPLEXITY AND CONNECTIVITY Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 [C] Gene Encodes a Plant-Specific Protein Required for Embryo Provasculature Development 1 [C] [W] Gene Encodes a Plant-Specific Protein Required for Embryo Provasculatur
    Article Snippet: Sections were then immersed in 100% ethanol for 2 min, air dried, prehybridized in hybridization buffer (50% [v/v] formamide, 1× SSC, 1× Denhardt’s solution [Amresco], 50 m m EDTA, 500 μg mL−1 tRNA, and 50 m m HEPES, pH 7) at 37°C for 30 min, and then hybridized with sense or antisense RNA probes overnight at 37°C in hybridization buffer. .. For signal detection, a 1:500-fold diluted primary streptavidin-horseradish peroxidase (0.5 mg mL−1 ; Thermo Scientific) solution was applied for 30 min on each section at room temperature.

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    Thermo Fisher poly hrp streptavidin
    IEC-6 cell lysate was incubated with biotin-CLEFMA (10 and 20 μM) or CLEFMA-N-biotin (10 μM) and separated on a gel. After transferring to a membrane, the proteins were probed <t>HRP-streptavidin.</t>
    Poly Hrp 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
    https://www.bioz.com/result/poly hrp streptavidin/product/Thermo Fisher
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    poly hrp streptavidin - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher streptavidin hrp
    Schematic diagram showed non-radioactive metabolic incorporation followed by azide-biotin or azide-Alex555 labeling, and biotin signals of proteins were detected by <t>streptavidin-HRP</t> by western blot. HPG is incorporated into newly synthesized proteins by metabolism and protein synthesis and the triazole conjugation between newly alkyne proteins labeled HPG and azide labeled either biotin or Alex555 via CuSO4 catalysis (A) . (B-a) The detection of biotin signals from extracted total proteins labeled by labeling reaction. Normal culture medium was changed to replace DMEM free of L-methionine supplemented with HPG after pulse 4 hr, and proteins were extracted in each of group at various time points including 0, 4, 24 and 72 hr. (B-b) Biotin signals of total proteins were detected. 1: Normal culture condition group; 2: HPG plus anisomycin group; 3: HPG group. (B-c,d,e) Biotin signals of Bcl-2, MMP-9 and IgG were individually detected in the immunoprecipitate pulled down by primary antibodies via siRNA post-transfection followed by non-radioactive metabolic labeling. (B-f) Radioactive isotope 35 S-methonine incorporated into synthesized IgG purified by immunoprecipitation was detected by autoradiography. 1: 35 S-methonine treated human choriocarcinoma cell line BeWo group and then antibody against human IgG immunoprecipitated human IgG in extracted total proteins; 2: cycloheximide plus 35 S-methonine treated BeWo group then antibody against human IgG immunoprecipitated human IgG in extracted total proteins; 3: 35 S-methonine treated skin fibroblast and then antibody against human IgG immunoprecipitated human IgG in extracted total proteins.
    Streptavidin Hrp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 162 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin hrp/product/Thermo Fisher
    Average 99 stars, based on 162 article reviews
    Price from $9.99 to $1999.99
    streptavidin hrp - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher horseradish peroxidase conjugated streptavidin
    Schematic diagram showed non-radioactive metabolic incorporation followed by azide-biotin or azide-Alex555 labeling, and biotin signals of proteins were detected by <t>streptavidin-HRP</t> by western blot. HPG is incorporated into newly synthesized proteins by metabolism and protein synthesis and the triazole conjugation between newly alkyne proteins labeled HPG and azide labeled either biotin or Alex555 via CuSO4 catalysis (A) . (B-a) The detection of biotin signals from extracted total proteins labeled by labeling reaction. Normal culture medium was changed to replace DMEM free of L-methionine supplemented with HPG after pulse 4 hr, and proteins were extracted in each of group at various time points including 0, 4, 24 and 72 hr. (B-b) Biotin signals of total proteins were detected. 1: Normal culture condition group; 2: HPG plus anisomycin group; 3: HPG group. (B-c,d,e) Biotin signals of Bcl-2, MMP-9 and IgG were individually detected in the immunoprecipitate pulled down by primary antibodies via siRNA post-transfection followed by non-radioactive metabolic labeling. (B-f) Radioactive isotope 35 S-methonine incorporated into synthesized IgG purified by immunoprecipitation was detected by autoradiography. 1: 35 S-methonine treated human choriocarcinoma cell line BeWo group and then antibody against human IgG immunoprecipitated human IgG in extracted total proteins; 2: cycloheximide plus 35 S-methonine treated BeWo group then antibody against human IgG immunoprecipitated human IgG in extracted total proteins; 3: 35 S-methonine treated skin fibroblast and then antibody against human IgG immunoprecipitated human IgG in extracted total proteins.
    Horseradish Peroxidase Conjugated Streptavidin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/horseradish peroxidase conjugated streptavidin/product/Thermo Fisher
    Average 99 stars, based on 90 article reviews
    Price from $9.99 to $1999.99
    horseradish peroxidase conjugated streptavidin - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    Image Search Results


    IEC-6 cell lysate was incubated with biotin-CLEFMA (10 and 20 μM) or CLEFMA-N-biotin (10 μM) and separated on a gel. After transferring to a membrane, the proteins were probed HRP-streptavidin.

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: IEC-6 cell lysate was incubated with biotin-CLEFMA (10 and 20 μM) or CLEFMA-N-biotin (10 μM) and separated on a gel. After transferring to a membrane, the proteins were probed HRP-streptavidin.

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Incubation, Transferring

    Effect of CLEFMA on proteolytic stability of RPN13. Whole cell lysate of H441 cells was treated with biotin-CLEFMA, followed by thermolysin digestion at room temperature for 10–60 min. The mixture was separated on an acrylamide gel, and the separated proteins were subjected to (A) Coomassie Brilliant staining, (B) HRP-Streptavidin probing for chemiluminescence imaging, and (C) Anti-RPN13 immunoblotting.

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: Effect of CLEFMA on proteolytic stability of RPN13. Whole cell lysate of H441 cells was treated with biotin-CLEFMA, followed by thermolysin digestion at room temperature for 10–60 min. The mixture was separated on an acrylamide gel, and the separated proteins were subjected to (A) Coomassie Brilliant staining, (B) HRP-Streptavidin probing for chemiluminescence imaging, and (C) Anti-RPN13 immunoblotting.

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Acrylamide Gel Assay, Staining, Imaging

    CLEFMA and EF24 interact with RPN13 in the fully assembled 26S proteasome and whole cell lysate. (A) The biotinylated compounds were allowed to react with purified human 26S proteasome, the mixture was separated on poly-acrylamide gels, and the transfer membrane was probed with HRP-streptavidin (blot #1). The blot was stripped and re-probed with anti-human RPN13 (blot #2). (B) Streptavidin bead-mediated pull down of RPN13 from rat IEC-6 cell lysate treated with biotinylated EF24 or CLEFMA. After blotting with anti-human HRP-streptavidin (blot #3), the membrane was stripped and re-probed with anti-rat RPN13 antibody (blot #4).

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: CLEFMA and EF24 interact with RPN13 in the fully assembled 26S proteasome and whole cell lysate. (A) The biotinylated compounds were allowed to react with purified human 26S proteasome, the mixture was separated on poly-acrylamide gels, and the transfer membrane was probed with HRP-streptavidin (blot #1). The blot was stripped and re-probed with anti-human RPN13 (blot #2). (B) Streptavidin bead-mediated pull down of RPN13 from rat IEC-6 cell lysate treated with biotinylated EF24 or CLEFMA. After blotting with anti-human HRP-streptavidin (blot #3), the membrane was stripped and re-probed with anti-rat RPN13 antibody (blot #4).

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Purification

    Interaction of EF24 and CLEFMA with proteasome. Highly purified (A,B) 26S proteasome, (C) 20S proteasome, and (D,E) 19S regulator were allowed to interact with biotinylated analogs of EF24 or CLEFMA (10 μM). The mixtures were separated on a regular 10% reducing gel and the transfer membranes were probed for biotin using HRP-streptavidin.

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: Interaction of EF24 and CLEFMA with proteasome. Highly purified (A,B) 26S proteasome, (C) 20S proteasome, and (D,E) 19S regulator were allowed to interact with biotinylated analogs of EF24 or CLEFMA (10 μM). The mixtures were separated on a regular 10% reducing gel and the transfer membranes were probed for biotin using HRP-streptavidin.

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Purification

    CLEFMA and EF24 interact with RPN13. Purified 19S regulatory particle was allowed to react with biotin-CLEFMA and the reaction mixture was separated by 2D-gel electrophoresis. (a) Coomassie-stained 2D-gel and (b) Corresponding transfer membrane probed with HRP-streptavidin. The four spots identified in the pictures were subjected to mass spectroscopic analysis (Table S1 ). (c) Recombinant human RPN13-Pru protein was reacted with various biotin-EF24 in presence (lane 6) or absence (lane 3) of free CLEFMA. Inactive biotinylated analogs of EF24 were also reacted as controls.

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: CLEFMA and EF24 interact with RPN13. Purified 19S regulatory particle was allowed to react with biotin-CLEFMA and the reaction mixture was separated by 2D-gel electrophoresis. (a) Coomassie-stained 2D-gel and (b) Corresponding transfer membrane probed with HRP-streptavidin. The four spots identified in the pictures were subjected to mass spectroscopic analysis (Table S1 ). (c) Recombinant human RPN13-Pru protein was reacted with various biotin-EF24 in presence (lane 6) or absence (lane 3) of free CLEFMA. Inactive biotinylated analogs of EF24 were also reacted as controls.

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Purification, Two-Dimensional Gel Electrophoresis, Electrophoresis, Staining, Recombinant

    IEC-6 cell lysate was incubated with biotin-CLEFMA (10 and 20 μM) or CLEFMA-N-biotin (10 μM) and separated on a gel. After transferring to a membrane, the proteins were probed HRP-streptavidin.

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: IEC-6 cell lysate was incubated with biotin-CLEFMA (10 and 20 μM) or CLEFMA-N-biotin (10 μM) and separated on a gel. After transferring to a membrane, the proteins were probed HRP-streptavidin.

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Incubation, Transferring

    Effect of CLEFMA on proteolytic stability of RPN13. Whole cell lysate of H441 cells was treated with biotin-CLEFMA, followed by thermolysin digestion at room temperature for 10–60 min. The mixture was separated on an acrylamide gel, and the separated proteins were subjected to (A) Coomassie Brilliant staining, (B) HRP-Streptavidin probing for chemiluminescence imaging, and (C) Anti-RPN13 immunoblotting.

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: Effect of CLEFMA on proteolytic stability of RPN13. Whole cell lysate of H441 cells was treated with biotin-CLEFMA, followed by thermolysin digestion at room temperature for 10–60 min. The mixture was separated on an acrylamide gel, and the separated proteins were subjected to (A) Coomassie Brilliant staining, (B) HRP-Streptavidin probing for chemiluminescence imaging, and (C) Anti-RPN13 immunoblotting.

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Acrylamide Gel Assay, Staining, Imaging

    CLEFMA and EF24 interact with RPN13 in the fully assembled 26S proteasome and whole cell lysate. (A) The biotinylated compounds were allowed to react with purified human 26S proteasome, the mixture was separated on poly-acrylamide gels, and the transfer membrane was probed with HRP-streptavidin (blot #1). The blot was stripped and re-probed with anti-human RPN13 (blot #2). (B) Streptavidin bead-mediated pull down of RPN13 from rat IEC-6 cell lysate treated with biotinylated EF24 or CLEFMA. After blotting with anti-human HRP-streptavidin (blot #3), the membrane was stripped and re-probed with anti-rat RPN13 antibody (blot #4).

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: CLEFMA and EF24 interact with RPN13 in the fully assembled 26S proteasome and whole cell lysate. (A) The biotinylated compounds were allowed to react with purified human 26S proteasome, the mixture was separated on poly-acrylamide gels, and the transfer membrane was probed with HRP-streptavidin (blot #1). The blot was stripped and re-probed with anti-human RPN13 (blot #2). (B) Streptavidin bead-mediated pull down of RPN13 from rat IEC-6 cell lysate treated with biotinylated EF24 or CLEFMA. After blotting with anti-human HRP-streptavidin (blot #3), the membrane was stripped and re-probed with anti-rat RPN13 antibody (blot #4).

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Purification

    Interaction of EF24 and CLEFMA with proteasome. Highly purified (A,B) 26S proteasome, (C) 20S proteasome, and (D,E) 19S regulator were allowed to interact with biotinylated analogs of EF24 or CLEFMA (10 μM). The mixtures were separated on a regular 10% reducing gel and the transfer membranes were probed for biotin using HRP-streptavidin.

    Journal: Frontiers in Chemistry

    Article Title: Ubiquitin Receptor RPN13 Mediates the Inhibitory Interaction of Diphenyldihaloketones CLEFMA and EF24 With the 26S Proteasome

    doi: 10.3389/fchem.2018.00392

    Figure Lengend Snippet: Interaction of EF24 and CLEFMA with proteasome. Highly purified (A,B) 26S proteasome, (C) 20S proteasome, and (D,E) 19S regulator were allowed to interact with biotinylated analogs of EF24 or CLEFMA (10 μM). The mixtures were separated on a regular 10% reducing gel and the transfer membranes were probed for biotin using HRP-streptavidin.

    Article Snippet: The membrane was blocked for 2 h in 5% Bovine Serum Albumin (BSA) in TTBS, before 2 h incubation in primary solution of poly-HRP Streptavidin [ThermoFisher Sci., Waltham, MA diluted 1:500,000 in 2% BSA in TTBS].

    Techniques: Purification

    Schematic diagram showed non-radioactive metabolic incorporation followed by azide-biotin or azide-Alex555 labeling, and biotin signals of proteins were detected by streptavidin-HRP by western blot. HPG is incorporated into newly synthesized proteins by metabolism and protein synthesis and the triazole conjugation between newly alkyne proteins labeled HPG and azide labeled either biotin or Alex555 via CuSO4 catalysis (A) . (B-a) The detection of biotin signals from extracted total proteins labeled by labeling reaction. Normal culture medium was changed to replace DMEM free of L-methionine supplemented with HPG after pulse 4 hr, and proteins were extracted in each of group at various time points including 0, 4, 24 and 72 hr. (B-b) Biotin signals of total proteins were detected. 1: Normal culture condition group; 2: HPG plus anisomycin group; 3: HPG group. (B-c,d,e) Biotin signals of Bcl-2, MMP-9 and IgG were individually detected in the immunoprecipitate pulled down by primary antibodies via siRNA post-transfection followed by non-radioactive metabolic labeling. (B-f) Radioactive isotope 35 S-methonine incorporated into synthesized IgG purified by immunoprecipitation was detected by autoradiography. 1: 35 S-methonine treated human choriocarcinoma cell line BeWo group and then antibody against human IgG immunoprecipitated human IgG in extracted total proteins; 2: cycloheximide plus 35 S-methonine treated BeWo group then antibody against human IgG immunoprecipitated human IgG in extracted total proteins; 3: 35 S-methonine treated skin fibroblast and then antibody against human IgG immunoprecipitated human IgG in extracted total proteins.

    Journal: BMC Cell Biology

    Article Title: A morphologic and semi-quantitative technique to analyze synthesis and release of specific proteins in cells

    doi: 10.1186/s12860-014-0045-1

    Figure Lengend Snippet: Schematic diagram showed non-radioactive metabolic incorporation followed by azide-biotin or azide-Alex555 labeling, and biotin signals of proteins were detected by streptavidin-HRP by western blot. HPG is incorporated into newly synthesized proteins by metabolism and protein synthesis and the triazole conjugation between newly alkyne proteins labeled HPG and azide labeled either biotin or Alex555 via CuSO4 catalysis (A) . (B-a) The detection of biotin signals from extracted total proteins labeled by labeling reaction. Normal culture medium was changed to replace DMEM free of L-methionine supplemented with HPG after pulse 4 hr, and proteins were extracted in each of group at various time points including 0, 4, 24 and 72 hr. (B-b) Biotin signals of total proteins were detected. 1: Normal culture condition group; 2: HPG plus anisomycin group; 3: HPG group. (B-c,d,e) Biotin signals of Bcl-2, MMP-9 and IgG were individually detected in the immunoprecipitate pulled down by primary antibodies via siRNA post-transfection followed by non-radioactive metabolic labeling. (B-f) Radioactive isotope 35 S-methonine incorporated into synthesized IgG purified by immunoprecipitation was detected by autoradiography. 1: 35 S-methonine treated human choriocarcinoma cell line BeWo group and then antibody against human IgG immunoprecipitated human IgG in extracted total proteins; 2: cycloheximide plus 35 S-methonine treated BeWo group then antibody against human IgG immunoprecipitated human IgG in extracted total proteins; 3: 35 S-methonine treated skin fibroblast and then antibody against human IgG immunoprecipitated human IgG in extracted total proteins.

    Article Snippet: Each diluted standard was incubated with primary antibody coated on the well and streptavidin-HRP and OD value for each sample was detected by ELISA.

    Techniques: Labeling, Western Blot, Synthesized, Conjugation Assay, Transfection, Purification, Immunoprecipitation, Autoradiography