streptavidin poly hrp  (Thermo Fisher)


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    Structured Review

    Thermo Fisher streptavidin poly hrp
    Oligonucleotides identified by ADAPT reveal aptamer-like characteristics. ( a ) Filter retention analysis of C1Q-binding by the ssODNs H1 (dark grey columns) and H11 (pale grey columns) at indicated C1Q concentrations. As a control, the reverse complement of H1, H1RC, was used (black columns). ( b ) ELONA analysis of C1Q-binding by the ssODNs H11 (circles) at indicated concentrations and a fixed C1Q concentration (0.625 nM). As a control, the reverse complement of H11, H11RC, was used (squares). “No aptamer” control (triangles) shows low background binding of detector <t>Streptavidin-HRP.</t> H11 specifically binds C1Q (estimated K D around 40 nM). ( c ) PAGE analysis of PEG-precipitated and ssODN-associated proteins pulled-down with L2. Lane 1: Molecular weight marker; lane 2: Input library L2; lane 3: Fraction pulled down by biotinylated L2; lane 4: Fraction pulled down by non-biotinylated L2; lane 5: Fraction found in the absence of DNA library; red arrows indicate the ssODN library; yellow arrows indicate protein bands cut out and analysed by LC-MS/MS; black arrows indicate streptavidin monomers leaking from beads. ( d ) Four-way Venn diagram of proteins detected by LC-MS/MS from Unfractionated plasma, PEG-precipitated plasma, PEG-precipitated plasma in presence of L0 or L2, respectively, purified by streptavidin magnetic beads (background-subtracted; i.e. biotinylated minus non-biotinylated libraries). ( e ) Gene ontology (GO) cellular component enrichment analysis of the subset of proteins associated with L0 (red bars) and L2 (grey bars) that show a p value of at least 6 × 10 −12 . Proteins listed are: 1 extracellular region part, 2 extracellular region, 3 extracellular exosome, 4 extracellular membrane-bounded organelle, 5 extracellular organelle, 6 extracellular vesicle, 7 membrane-bounded vesicle, 8 vesicle, 9 organelle, 10 membrane-bounded organelle, 11 extracellular space, 12 focal adhesion, 13 cell-substrate adherence junction, 14 cell-substrate junction, 15 adherence junction, 16 anchoring junction, 17 cellular component, 18 cell junction, 19 blood micro particle. Inset: 108 proteins pulled down by L2 (inset, 96 + 12), cut from the gel shown in ( c ), and analysed by LC-MS/MS, 13 proteins unique to background-subtracted L0 (inset, 13), and 12 overlapping proteins (inset, 12).
    Streptavidin Poly Hrp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin poly hrp/product/Thermo Fisher
    Average 99 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    streptavidin poly hrp - by Bioz Stars, 2020-03
    99/100 stars

    Images

    1) Product Images from "Plasma Exosome Profiling of Cancer Patients by a Next Generation Systems Biology Approach"

    Article Title: Plasma Exosome Profiling of Cancer Patients by a Next Generation Systems Biology Approach

    Journal: Scientific Reports

    doi: 10.1038/srep42741

    Oligonucleotides identified by ADAPT reveal aptamer-like characteristics. ( a ) Filter retention analysis of C1Q-binding by the ssODNs H1 (dark grey columns) and H11 (pale grey columns) at indicated C1Q concentrations. As a control, the reverse complement of H1, H1RC, was used (black columns). ( b ) ELONA analysis of C1Q-binding by the ssODNs H11 (circles) at indicated concentrations and a fixed C1Q concentration (0.625 nM). As a control, the reverse complement of H11, H11RC, was used (squares). “No aptamer” control (triangles) shows low background binding of detector Streptavidin-HRP. H11 specifically binds C1Q (estimated K D around 40 nM). ( c ) PAGE analysis of PEG-precipitated and ssODN-associated proteins pulled-down with L2. Lane 1: Molecular weight marker; lane 2: Input library L2; lane 3: Fraction pulled down by biotinylated L2; lane 4: Fraction pulled down by non-biotinylated L2; lane 5: Fraction found in the absence of DNA library; red arrows indicate the ssODN library; yellow arrows indicate protein bands cut out and analysed by LC-MS/MS; black arrows indicate streptavidin monomers leaking from beads. ( d ) Four-way Venn diagram of proteins detected by LC-MS/MS from Unfractionated plasma, PEG-precipitated plasma, PEG-precipitated plasma in presence of L0 or L2, respectively, purified by streptavidin magnetic beads (background-subtracted; i.e. biotinylated minus non-biotinylated libraries). ( e ) Gene ontology (GO) cellular component enrichment analysis of the subset of proteins associated with L0 (red bars) and L2 (grey bars) that show a p value of at least 6 × 10 −12 . Proteins listed are: 1 extracellular region part, 2 extracellular region, 3 extracellular exosome, 4 extracellular membrane-bounded organelle, 5 extracellular organelle, 6 extracellular vesicle, 7 membrane-bounded vesicle, 8 vesicle, 9 organelle, 10 membrane-bounded organelle, 11 extracellular space, 12 focal adhesion, 13 cell-substrate adherence junction, 14 cell-substrate junction, 15 adherence junction, 16 anchoring junction, 17 cellular component, 18 cell junction, 19 blood micro particle. Inset: 108 proteins pulled down by L2 (inset, 96 + 12), cut from the gel shown in ( c ), and analysed by LC-MS/MS, 13 proteins unique to background-subtracted L0 (inset, 13), and 12 overlapping proteins (inset, 12).
    Figure Legend Snippet: Oligonucleotides identified by ADAPT reveal aptamer-like characteristics. ( a ) Filter retention analysis of C1Q-binding by the ssODNs H1 (dark grey columns) and H11 (pale grey columns) at indicated C1Q concentrations. As a control, the reverse complement of H1, H1RC, was used (black columns). ( b ) ELONA analysis of C1Q-binding by the ssODNs H11 (circles) at indicated concentrations and a fixed C1Q concentration (0.625 nM). As a control, the reverse complement of H11, H11RC, was used (squares). “No aptamer” control (triangles) shows low background binding of detector Streptavidin-HRP. H11 specifically binds C1Q (estimated K D around 40 nM). ( c ) PAGE analysis of PEG-precipitated and ssODN-associated proteins pulled-down with L2. Lane 1: Molecular weight marker; lane 2: Input library L2; lane 3: Fraction pulled down by biotinylated L2; lane 4: Fraction pulled down by non-biotinylated L2; lane 5: Fraction found in the absence of DNA library; red arrows indicate the ssODN library; yellow arrows indicate protein bands cut out and analysed by LC-MS/MS; black arrows indicate streptavidin monomers leaking from beads. ( d ) Four-way Venn diagram of proteins detected by LC-MS/MS from Unfractionated plasma, PEG-precipitated plasma, PEG-precipitated plasma in presence of L0 or L2, respectively, purified by streptavidin magnetic beads (background-subtracted; i.e. biotinylated minus non-biotinylated libraries). ( e ) Gene ontology (GO) cellular component enrichment analysis of the subset of proteins associated with L0 (red bars) and L2 (grey bars) that show a p value of at least 6 × 10 −12 . Proteins listed are: 1 extracellular region part, 2 extracellular region, 3 extracellular exosome, 4 extracellular membrane-bounded organelle, 5 extracellular organelle, 6 extracellular vesicle, 7 membrane-bounded vesicle, 8 vesicle, 9 organelle, 10 membrane-bounded organelle, 11 extracellular space, 12 focal adhesion, 13 cell-substrate adherence junction, 14 cell-substrate junction, 15 adherence junction, 16 anchoring junction, 17 cellular component, 18 cell junction, 19 blood micro particle. Inset: 108 proteins pulled down by L2 (inset, 96 + 12), cut from the gel shown in ( c ), and analysed by LC-MS/MS, 13 proteins unique to background-subtracted L0 (inset, 13), and 12 overlapping proteins (inset, 12).

    Techniques Used: Binding Assay, Concentration Assay, Polyacrylamide Gel Electrophoresis, Molecular Weight, Marker, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Purification, Magnetic Beads

    2) Product Images from "Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †"

    Article Title: Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †

    Journal: Lab on a Chip

    doi: 10.1039/c1lc20833k

    Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.
    Figure Legend Snippet: Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.

    Techniques Used: Amplification

    3) Product Images from "Point-of-care test for cervical cancer in LMICs"

    Article Title: Point-of-care test for cervical cancer in LMICs

    Journal: Oncotarget

    doi: 10.18632/oncotarget.7709

    Scheme of enzyme-enhanced LFIC for VCP detection Biotinylated gold nanoparticles tethered with streptavidin-bearing HRP at the capture site in the lateral flow strip to generate a signal upon interaction with TMB.
    Figure Legend Snippet: Scheme of enzyme-enhanced LFIC for VCP detection Biotinylated gold nanoparticles tethered with streptavidin-bearing HRP at the capture site in the lateral flow strip to generate a signal upon interaction with TMB.

    Techniques Used: Flow Cytometry, Stripping Membranes

    4) Product Images from "Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †"

    Article Title: Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †

    Journal: Lab on a Chip

    doi: 10.1039/c1lc20833k

    Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.
    Figure Legend Snippet: Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.

    Techniques Used: Amplification

    5) Product Images from "Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †"

    Article Title: Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †

    Journal: Lab on a Chip

    doi: 10.1039/c1lc20833k

    Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.
    Figure Legend Snippet: Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.

    Techniques Used: Amplification

    6) Product Images from "Med5(Nut1) and Med17(Srb4) Are Direct Targets of Mediator Histone H4 Tail Interactions"

    Article Title: Med5(Nut1) and Med17(Srb4) Are Direct Targets of Mediator Histone H4 Tail Interactions

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0038416

    H4 10 Bpa cross-linking patterns of the mutant Mediator complexes. SDS-PAGE blot probed with streptavidin poly-HRP to detect and compare the pattern of biotinylated peptide cross-linked to Mediator subunits in WT Mediator and the mutant Mediator complexes. Equimolar amounts (∼7.5 nM) of each indicated Mediator complex were incubated with H4 10 Bpa (4 µM) and exposed to UV for 15 min. Asterisk refers to the same weak signal as in Fig. 2-A.
    Figure Legend Snippet: H4 10 Bpa cross-linking patterns of the mutant Mediator complexes. SDS-PAGE blot probed with streptavidin poly-HRP to detect and compare the pattern of biotinylated peptide cross-linked to Mediator subunits in WT Mediator and the mutant Mediator complexes. Equimolar amounts (∼7.5 nM) of each indicated Mediator complex were incubated with H4 10 Bpa (4 µM) and exposed to UV for 15 min. Asterisk refers to the same weak signal as in Fig. 2-A.

    Techniques Used: Mutagenesis, SDS Page, Incubation

    H4 10 Bpa has four specific cross-linking targets within Mediator. (A) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated peptide cross-linked to Mediator subunits. WT H4, H4 10 Bpa and H4 22 Bpa (4 µM) were incubated in the presence or absence of the WT Mediator complex (∼7.5 nM) and exposed to UV for 15 min when indicated. Cross-linking products were resolved on 6% SDS-polyacrylamide gel, transferred to to PVDF, detected by streptavidin poly-HRP, and referred to as BCTs (H4 10 B pa C ross-linking T argets). A weak band with relatively poor reproducibility was asterisked. (B) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits after different UV exposure times. Identical mixtures, which contain 7.5 nM Mediator complex and 4 µM H4 10 Bpa, were exposed to UV for 0, 5, 10 or 15 min. (C) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits in reactions with varient H4 10 Bpa concentration. WT Mediator complex (∼7.5 nM) was incubated with 1 µM, 2 µM or 4 µM H4 10 Bpa peptide and exposed to UV irradiation for 15 min. (D) Western blot analysis of histone tail peptide binding experiment comparing Mediator binding affinity for H4 10 Bpa and H2B tail peptide under the identical concentrations to the cross-linking reactions. WT Mediator complex (∼3 nM) was mixed with H4 10 Bpa (4 µM) or synthetic biotinylated histone H2B N’-tail peptide (4 µM or 8 µM) as the inputs. The basic steps and layout of the analysis were as described earlier ( Fig. 1 ). (E) and (F) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits, after H4 22 Bpa (E) or H2B tail peptide (F) was added at the indicated concentration.
    Figure Legend Snippet: H4 10 Bpa has four specific cross-linking targets within Mediator. (A) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated peptide cross-linked to Mediator subunits. WT H4, H4 10 Bpa and H4 22 Bpa (4 µM) were incubated in the presence or absence of the WT Mediator complex (∼7.5 nM) and exposed to UV for 15 min when indicated. Cross-linking products were resolved on 6% SDS-polyacrylamide gel, transferred to to PVDF, detected by streptavidin poly-HRP, and referred to as BCTs (H4 10 B pa C ross-linking T argets). A weak band with relatively poor reproducibility was asterisked. (B) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits after different UV exposure times. Identical mixtures, which contain 7.5 nM Mediator complex and 4 µM H4 10 Bpa, were exposed to UV for 0, 5, 10 or 15 min. (C) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits in reactions with varient H4 10 Bpa concentration. WT Mediator complex (∼7.5 nM) was incubated with 1 µM, 2 µM or 4 µM H4 10 Bpa peptide and exposed to UV irradiation for 15 min. (D) Western blot analysis of histone tail peptide binding experiment comparing Mediator binding affinity for H4 10 Bpa and H2B tail peptide under the identical concentrations to the cross-linking reactions. WT Mediator complex (∼3 nM) was mixed with H4 10 Bpa (4 µM) or synthetic biotinylated histone H2B N’-tail peptide (4 µM or 8 µM) as the inputs. The basic steps and layout of the analysis were as described earlier ( Fig. 1 ). (E) and (F) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits, after H4 22 Bpa (E) or H2B tail peptide (F) was added at the indicated concentration.

    Techniques Used: SDS Page, Incubation, Concentration Assay, Irradiation, Western Blot, Binding Assay

    Med5(Nut1)p, Med14(Rgr1)p, Med17(Srb4)p and Med1p are H4 10 Bpa cross-linking targets. Silver staining (A) and immunoblotting analysis (B) comparing the composition of affinity-purified MYC -tagged and non- MYC -tagged WT Mediator complexes after 10% SDS-PAGE. (C) A comparison of the cross-linking patterns of MYC -tagged Mediator complexes with the WT pattern using an SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits. Each indicated Mediator species (∼7.5 nM) was incubated with H4 10 Bpa (4 µM). A ‘Long Exposure’ of the 79 kD region on the SDS-PAGE blot probed with streptavidin poly-HRP is shown for a better view of the weak BCT4 signal in each sample.
    Figure Legend Snippet: Med5(Nut1)p, Med14(Rgr1)p, Med17(Srb4)p and Med1p are H4 10 Bpa cross-linking targets. Silver staining (A) and immunoblotting analysis (B) comparing the composition of affinity-purified MYC -tagged and non- MYC -tagged WT Mediator complexes after 10% SDS-PAGE. (C) A comparison of the cross-linking patterns of MYC -tagged Mediator complexes with the WT pattern using an SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits. Each indicated Mediator species (∼7.5 nM) was incubated with H4 10 Bpa (4 µM). A ‘Long Exposure’ of the 79 kD region on the SDS-PAGE blot probed with streptavidin poly-HRP is shown for a better view of the weak BCT4 signal in each sample.

    Techniques Used: Silver Staining, Affinity Purification, SDS Page, Incubation

    7) Product Images from "N-linked glycosylation modulates the immunogenicity of recombinant human factor VIII in hemophilia A mice"

    Article Title: N-linked glycosylation modulates the immunogenicity of recombinant human factor VIII in hemophilia A mice

    Journal: Haematologica

    doi: 10.3324/haematol.2018.188219

    Lectin array analysis of exposed glycans on rFVIII products. rFVIII products were adsorbed on microtitre plates at 1 μg/mL and assessed using a panel of biotinylated lectins. BSA was adsorbed as a control. Binding was detected using a streptavidin poly-HRP and read at 492 nm. (A) Heat plot demonstrating the median-centred binding values of lectins with varying carbohydrate specificites to different rFVIII products. Values shown are representative of the mean of at least 3 independent experiments on a single lot of each rFVIII product. Statistical summary available in Online Supplementary Table S1 . (B) Lectin binding analysis of different production lots of full-length rFVIII from CHO or BHK cells. Data are representative of at least 3 independent experiments. BHK: baby hamster kidney cells; CHO: Chinese hamster ovary cells: rFVIII: recombinant factor VIII; BSA: bovine serum albumin; BDD: B-domain deleted.
    Figure Legend Snippet: Lectin array analysis of exposed glycans on rFVIII products. rFVIII products were adsorbed on microtitre plates at 1 μg/mL and assessed using a panel of biotinylated lectins. BSA was adsorbed as a control. Binding was detected using a streptavidin poly-HRP and read at 492 nm. (A) Heat plot demonstrating the median-centred binding values of lectins with varying carbohydrate specificites to different rFVIII products. Values shown are representative of the mean of at least 3 independent experiments on a single lot of each rFVIII product. Statistical summary available in Online Supplementary Table S1 . (B) Lectin binding analysis of different production lots of full-length rFVIII from CHO or BHK cells. Data are representative of at least 3 independent experiments. BHK: baby hamster kidney cells; CHO: Chinese hamster ovary cells: rFVIII: recombinant factor VIII; BSA: bovine serum albumin; BDD: B-domain deleted.

    Techniques Used: Binding Assay, Recombinant

    8) Product Images from "Regulation of the Edwardsiella ictaluri Type III Secretion System by pH and Phosphate Concentration through EsrA, EsrB, and EsrC ▿"

    Article Title: Regulation of the Edwardsiella ictaluri Type III Secretion System by pH and Phosphate Concentration through EsrA, EsrB, and EsrC ▿

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.00195-11

    Western blot detection of T3SS proteins fused to the Flag epitope in Edwardsiella ictaluri whole-cell lysates. Strains carrying the fusions were cultured in MM19 at pH 7.0, MM19 at pH 5.5, and MMP at pH 5.5. Proteins carrying the Flag epitope were labeled with anti-Flag antibody followed by HRP-conjugated goat anti-mouse antibody to detect EsrC, EseB, and EseG. Detection of EseI required mouse anti-Flag antibody followed by biotinylated goat anti-mouse antibody and HRP-conjugated streptavidin. Loading of equivalent protein amounts across lanes was verified using mouse anti-DnaK antibody, biotinylated goat anti-mouse antibody, and HRP-conjugated streptavidin.
    Figure Legend Snippet: Western blot detection of T3SS proteins fused to the Flag epitope in Edwardsiella ictaluri whole-cell lysates. Strains carrying the fusions were cultured in MM19 at pH 7.0, MM19 at pH 5.5, and MMP at pH 5.5. Proteins carrying the Flag epitope were labeled with anti-Flag antibody followed by HRP-conjugated goat anti-mouse antibody to detect EsrC, EseB, and EseG. Detection of EseI required mouse anti-Flag antibody followed by biotinylated goat anti-mouse antibody and HRP-conjugated streptavidin. Loading of equivalent protein amounts across lanes was verified using mouse anti-DnaK antibody, biotinylated goat anti-mouse antibody, and HRP-conjugated streptavidin.

    Techniques Used: Western Blot, FLAG-tag, Cell Culture, Labeling

    9) Product Images from "Med5(Nut1) and Med17(Srb4) Are Direct Targets of Mediator Histone H4 Tail Interactions"

    Article Title: Med5(Nut1) and Med17(Srb4) Are Direct Targets of Mediator Histone H4 Tail Interactions

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0038416

    H4 10 Bpa cross-linking patterns of the mutant Mediator complexes. SDS-PAGE blot probed with streptavidin poly-HRP to detect and compare the pattern of biotinylated peptide cross-linked to Mediator subunits in WT Mediator and the mutant Mediator complexes. Equimolar amounts (∼7.5 nM) of each indicated Mediator complex were incubated with H4 10 Bpa (4 µM) and exposed to UV for 15 min. Asterisk refers to the same weak signal as in Fig. 2-A.
    Figure Legend Snippet: H4 10 Bpa cross-linking patterns of the mutant Mediator complexes. SDS-PAGE blot probed with streptavidin poly-HRP to detect and compare the pattern of biotinylated peptide cross-linked to Mediator subunits in WT Mediator and the mutant Mediator complexes. Equimolar amounts (∼7.5 nM) of each indicated Mediator complex were incubated with H4 10 Bpa (4 µM) and exposed to UV for 15 min. Asterisk refers to the same weak signal as in Fig. 2-A.

    Techniques Used: Mutagenesis, SDS Page, Incubation

    H4 10 Bpa has four specific cross-linking targets within Mediator. (A) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated peptide cross-linked to Mediator subunits. WT H4, H4 10 Bpa and H4 22 Bpa (4 µM) were incubated in the presence or absence of the WT Mediator complex (∼7.5 nM) and exposed to UV for 15 min when indicated. Cross-linking products were resolved on 6% SDS-polyacrylamide gel, transferred to to PVDF, detected by streptavidin poly-HRP, and referred to as BCTs (H4 10 B pa C ross-linking T argets). A weak band with relatively poor reproducibility was asterisked. (B) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits after different UV exposure times. Identical mixtures, which contain 7.5 nM Mediator complex and 4 µM H4 10 Bpa, were exposed to UV for 0, 5, 10 or 15 min. (C) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits in reactions with varient H4 10 Bpa concentration. WT Mediator complex (∼7.5 nM) was incubated with 1 µM, 2 µM or 4 µM H4 10 Bpa peptide and exposed to UV irradiation for 15 min. (D) Western blot analysis of histone tail peptide binding experiment comparing Mediator binding affinity for H4 10 Bpa and H2B tail peptide under the identical concentrations to the cross-linking reactions. WT Mediator complex (∼3 nM) was mixed with H4 10 Bpa (4 µM) or synthetic biotinylated histone H2B N’-tail peptide (4 µM or 8 µM) as the inputs. The basic steps and layout of the analysis were as described earlier ( Fig. 1 ). (E) and (F) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits, after H4 22 Bpa (E) or H2B tail peptide (F) was added at the indicated concentration.
    Figure Legend Snippet: H4 10 Bpa has four specific cross-linking targets within Mediator. (A) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated peptide cross-linked to Mediator subunits. WT H4, H4 10 Bpa and H4 22 Bpa (4 µM) were incubated in the presence or absence of the WT Mediator complex (∼7.5 nM) and exposed to UV for 15 min when indicated. Cross-linking products were resolved on 6% SDS-polyacrylamide gel, transferred to to PVDF, detected by streptavidin poly-HRP, and referred to as BCTs (H4 10 B pa C ross-linking T argets). A weak band with relatively poor reproducibility was asterisked. (B) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits after different UV exposure times. Identical mixtures, which contain 7.5 nM Mediator complex and 4 µM H4 10 Bpa, were exposed to UV for 0, 5, 10 or 15 min. (C) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits in reactions with varient H4 10 Bpa concentration. WT Mediator complex (∼7.5 nM) was incubated with 1 µM, 2 µM or 4 µM H4 10 Bpa peptide and exposed to UV irradiation for 15 min. (D) Western blot analysis of histone tail peptide binding experiment comparing Mediator binding affinity for H4 10 Bpa and H2B tail peptide under the identical concentrations to the cross-linking reactions. WT Mediator complex (∼3 nM) was mixed with H4 10 Bpa (4 µM) or synthetic biotinylated histone H2B N’-tail peptide (4 µM or 8 µM) as the inputs. The basic steps and layout of the analysis were as described earlier ( Fig. 1 ). (E) and (F) SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits, after H4 22 Bpa (E) or H2B tail peptide (F) was added at the indicated concentration.

    Techniques Used: SDS Page, Incubation, Concentration Assay, Irradiation, Western Blot, Binding Assay

    Med5(Nut1)p, Med14(Rgr1)p, Med17(Srb4)p and Med1p are H4 10 Bpa cross-linking targets. Silver staining (A) and immunoblotting analysis (B) comparing the composition of affinity-purified MYC -tagged and non- MYC -tagged WT Mediator complexes after 10% SDS-PAGE. (C) A comparison of the cross-linking patterns of MYC -tagged Mediator complexes with the WT pattern using an SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits. Each indicated Mediator species (∼7.5 nM) was incubated with H4 10 Bpa (4 µM). A ‘Long Exposure’ of the 79 kD region on the SDS-PAGE blot probed with streptavidin poly-HRP is shown for a better view of the weak BCT4 signal in each sample.
    Figure Legend Snippet: Med5(Nut1)p, Med14(Rgr1)p, Med17(Srb4)p and Med1p are H4 10 Bpa cross-linking targets. Silver staining (A) and immunoblotting analysis (B) comparing the composition of affinity-purified MYC -tagged and non- MYC -tagged WT Mediator complexes after 10% SDS-PAGE. (C) A comparison of the cross-linking patterns of MYC -tagged Mediator complexes with the WT pattern using an SDS-PAGE blot probed with streptavidin poly-HRP to detect biotinylated H4 10 Bpa peptide cross-linked to Mediator subunits. Each indicated Mediator species (∼7.5 nM) was incubated with H4 10 Bpa (4 µM). A ‘Long Exposure’ of the 79 kD region on the SDS-PAGE blot probed with streptavidin poly-HRP is shown for a better view of the weak BCT4 signal in each sample.

    Techniques Used: Silver Staining, Affinity Purification, SDS Page, Incubation

    10) Product Images from "Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †"

    Article Title: Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †

    Journal: Lab on a Chip

    doi: 10.1039/c1lc20833k

    Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.
    Figure Legend Snippet: Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.

    Techniques Used: Amplification

    11) Product Images from "Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †"

    Article Title: Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †

    Journal: Lab on a Chip

    doi: 10.1039/c1lc20833k

    Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.
    Figure Legend Snippet: Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.

    Techniques Used: Amplification

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    Article Snippet: Next, slides were incubated in 3% hydrogen peroxide in water for 10 minutes at room temperature, washed 4 times for 5 minutes in PBS with 0.1% Tween-20 (PBS-T), incubated with protein block (Leica, RE7102) for 30 minutes at room temperature, and then washed 4 times for 5 minutes in PBS-T. For enzymatic labeling, cells were incubated in a buffer containing 20 mM Tris-Cl pH 7.4, 50 mM DTT, 50 μM biotinylated NAD+ (Trevigen, 4670), with or without 20 ng/μL tagraxofusp at 37°C for 1 hour in a humid box. .. After incubation, slides were washed 4 times for 5 minutes each in PBS-T, and then incubated with streptavidin-poly HRP (Thermo Fisher Scientific, 21140) in PBS containing 1% BSA for 30 minutes at room temperature in a humid box.

    Article Title: BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells
    Article Snippet: The IgA1 and IgA2 recombinant proteins (16‐16‐090701‐1M and 16‐16‐090701‐2M, Athens Research) were serially diluted in blocking buffer to obtain a standard curve. .. Plates were washed 6 times and 1:10 000 streptavidin poly HRP (#21140, Thermo Fisher Scientific) was added and incubated for 1 h at room temperature while shaking (180 rpm).

    Article Title: ALPPL2 Is a Potential Diagnostic Biomarker for Pancreatic Cancer-Derived Extracellular Vesicles
    Article Snippet: After blocking, 20 pmol of SQ2 and SQ2 mutant aptamers (St Pharma, South Korea) in binding buffer (PBS with 5 mM MgCl2 , 0.5% BSA, 0.1 mg/mL yeast tRNA, 0.05% Tween-20) were added onto each well and kept for 1 h at RT. .. Wells were washed four times with washing buffer, and 2,000-fold diluted streptavidin-poly-HRP conjugate (Pierce) in 0.5% BSA-PBS was added and incubated for 30 min at RT.

    Enzyme-linked Immunosorbent Assay:

    Article Title: An ABCA1-independent pathway for recycling a poorly lipidated 8.1 nm apolipoprotein E particle from glia
    Article Snippet: .. Streptavidin-poly-HRP antibody (1:4000; Pierce) was then added and incubated at RT for 90 min. Color was developed by adding 100 μl/well Sigma ELISA TMB and stopped immediately by adding 100 μl of 1M HCL. .. Total cholesterol measurements of GCM samples were performed using the fluorogenic Amplex Red Cholesterol Assay Kit (Invitrogen) following the manufacture's protocol.

    Article Title: BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells
    Article Snippet: Paragraph title: IgA1 and IgA2 ELISA ... Plates were washed 6 times and 1:10 000 streptavidin poly HRP (#21140, Thermo Fisher Scientific) was added and incubated for 1 h at room temperature while shaking (180 rpm).

    Sandwich ELISA:

    Article Title: ALPPL2 Is a Potential Diagnostic Biomarker for Pancreatic Cancer-Derived Extracellular Vesicles
    Article Snippet: Paragraph title: Aptamer-Based Direct and Sandwich ELISA ... Wells were washed four times with washing buffer, and 2,000-fold diluted streptavidin-poly-HRP conjugate (Pierce) in 0.5% BSA-PBS was added and incubated for 30 min at RT.

    Incubation:

    Article Title: DNA methyltransferase inhibition overcomes diphthamide pathway deficiencies underlying CD123-targeted treatment resistance
    Article Snippet: .. After incubation, slides were washed 4 times for 5 minutes each in PBS-T, and then incubated with streptavidin-poly HRP (Thermo Fisher Scientific, 21140) in PBS containing 1% BSA for 30 minutes at room temperature in a humid box. .. Slides were washed 4 times for 5 minutes each in PBS-T and treated with DAB chromogen (Vector Labs, SK-4105) diluted in DAB diluent per the manufacturer’s instructions.

    Article Title: Lactosylceramide synthases encoded by B4galt5 and 6 genes are pivotal for neuronal generation and myelin formation in mice
    Article Snippet: The membranes were incubated overnight with primary antibodies in Can Get Signal immunoreaction enhancer solution (Toyobo, Osaka, Japan) at 4°C. .. Antibodies and dilutions used in western blotting were as follows: rabbit anti-TrkA Y490ph (1:500; CST, MA, USA, 9141s), rabbit anti-TrkA (1:500; Abcam, Cambridge, UK, 76291), rabbit anti-Lyn Y396ph (1:2000; Abcam, 40660), rabbit anti-Lyn (1:500; CST, 2796s), goat anti-rabbit IgG-HRP (1:2000; Abcam, 97023), goat anti-rabbit IgG-biotin (1:1000; Vector, CA, USA, BA-1000), and Pierce streptavidin poly-HRP (1:3000; Thermo Scientific, MA, USA, 21140).

    Article Title: An ABCA1-independent pathway for recycling a poorly lipidated 8.1 nm apolipoprotein E particle from glia
    Article Snippet: .. Streptavidin-poly-HRP antibody (1:4000; Pierce) was then added and incubated at RT for 90 min. Color was developed by adding 100 μl/well Sigma ELISA TMB and stopped immediately by adding 100 μl of 1M HCL. .. Total cholesterol measurements of GCM samples were performed using the fluorogenic Amplex Red Cholesterol Assay Kit (Invitrogen) following the manufacture's protocol.

    Article Title: SOD1-positive aggregate accumulation in the CNS predicts slower disease progression and increased longevity in a mutant SOD1 mouse model of ALS
    Article Snippet: .. After 1 h plates were washed and incubated for an additional hour with a 1:10,000 dilution of Streptavidin Poly-HRP (Pierce 21140). .. After the last wash, 50 μL of TMB HRP substrate solution (Surmodics) was added to each well and color development was stopped with 25 μL per well of 2 N H2SO4 (R & D DY994).

    Article Title: Expression of Components of the Renin-Angiotensin System in Pyogenic Granuloma
    Article Snippet: The samples were them probed using the primary antibodies ACE (1:200; cat# sc-12184, Santa Cruz, Rockford, IL, USA), PRR (1:500; cat# ab40790, Abcam, Cambridge, UK), ATIIR1 (1:200; cat# sc-1173, Santa Cruz), ATIIR2 (1:500; cat# ab92445, Abcam) and β-actin (1:500; cat# ab8229 Abcam); then incubated with the appropriate secondary antibody: goat anti-rabbit HRP (1:1000, cat# A16110, Thermo Fisher) and donkey anti-goat HRP (1:1,000, cat# ab97120, Abcam). .. ACE was probed using a tertiary cascade consisting of a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:4,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min.

    Article Title: BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells
    Article Snippet: .. Plates were washed 6 times and 1:10 000 streptavidin poly HRP (#21140, Thermo Fisher Scientific) was added and incubated for 1 h at room temperature while shaking (180 rpm). .. 100 μL TMB (#37574 Thermo Fisher Scientific) was added.

    Article Title: ALPPL2 Is a Potential Diagnostic Biomarker for Pancreatic Cancer-Derived Extracellular Vesicles
    Article Snippet: .. Wells were washed four times with washing buffer, and 2,000-fold diluted streptavidin-poly-HRP conjugate (Pierce) in 0.5% BSA-PBS was added and incubated for 30 min at RT. .. After four washes in wash buffer, ultra TMB-ELISA reagent (Thermo Scientific) was added and incubated for 5–15 min to allow blue color expression.

    Article Title: Utilizing Biotinylated Proteins Expressed in Yeast to Visualize DNA–Protein Interactions at the Single-Molecule Level
    Article Snippet: Boiled whole-protein extracts or purified protein samples were incubated with unboiled streptavidin for 5 min at room temperature and then loaded onto a sodium dodecyl sulfate-polyacrylamide gel. .. The antibodies used in this study were CBP tag antibody (A01798, Genscript), Flag tag antibody (F3165, Sigma-Aldrich), and streptavidin poly-HRP (21140, Thermo Fisher Scientific).

    Expressing:

    Article Title: ALPPL2 Is a Potential Diagnostic Biomarker for Pancreatic Cancer-Derived Extracellular Vesicles
    Article Snippet: Wells were washed four times with washing buffer, and 2,000-fold diluted streptavidin-poly-HRP conjugate (Pierce) in 0.5% BSA-PBS was added and incubated for 30 min at RT. .. After four washes in wash buffer, ultra TMB-ELISA reagent (Thermo Scientific) was added and incubated for 5–15 min to allow blue color expression.

    Western Blot:

    Article Title: Lactosylceramide synthases encoded by B4galt5 and 6 genes are pivotal for neuronal generation and myelin formation in mice
    Article Snippet: .. Antibodies and dilutions used in western blotting were as follows: rabbit anti-TrkA Y490ph (1:500; CST, MA, USA, 9141s), rabbit anti-TrkA (1:500; Abcam, Cambridge, UK, 76291), rabbit anti-Lyn Y396ph (1:2000; Abcam, 40660), rabbit anti-Lyn (1:500; CST, 2796s), goat anti-rabbit IgG-HRP (1:2000; Abcam, 97023), goat anti-rabbit IgG-biotin (1:1000; Vector, CA, USA, BA-1000), and Pierce streptavidin poly-HRP (1:3000; Thermo Scientific, MA, USA, 21140). .. Western blotting for brain homogenates was carried out, as described previously [ ], using total brain proteins, extracted using total protein extraction kit (P501S; 101 Bio, AC, USA).

    Article Title: Glioblastoma Multiforme Cancer Stem Cells Express Components of the Renin–Angiotensin System
    Article Snippet: Paragraph title: Western Blotting ... ACE tertiary cascade used a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:20,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min. β-actin antibody probing was performed with the iBind™ Flex device (cat# SLF2000, Life Technologies) using primary mouse monoclonal anti-β-actin (1:2000 cat# ab8226, Abcam) and secondary donkey anti-mouse Alexa fluor 488 (1:2000; cat# A21202, Thermo Fisher).

    Article Title: Expression of Components of the Renin-Angiotensin System in Pyogenic Granuloma
    Article Snippet: Paragraph title: Western Blotting ... ACE was probed using a tertiary cascade consisting of a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:4,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min.

    Derivative Assay:

    Article Title: SOD1-positive aggregate accumulation in the CNS predicts slower disease progression and increased longevity in a mutant SOD1 mouse model of ALS
    Article Snippet: A standard curve of denatured human erythrocyte derived SOD1 (see preparation of denatured SOD1 above) was included with serial two-fold dilutions from 0.3 to 0.0003 μg/mL as well as background control wells with assay buffer only. .. After 1 h plates were washed and incubated for an additional hour with a 1:10,000 dilution of Streptavidin Poly-HRP (Pierce 21140).

    Cell Culture:

    Article Title: Lactosylceramide synthases encoded by B4galt5 and 6 genes are pivotal for neuronal generation and myelin formation in mice
    Article Snippet: Western blot analysis Neurospheres cultured on 5 μg/mL laminin-coated glass chambers for differentiation were lysed in NP-40 buffer (150 mM NaCl, 50 mM Tris-Cl, 1% NP-40), containing a phosphatase inhibitor cocktail (Nacalai, Kyoto, Japan), and sonicated with Bioruptor (Diagenode, Liege, Belgium). .. Antibodies and dilutions used in western blotting were as follows: rabbit anti-TrkA Y490ph (1:500; CST, MA, USA, 9141s), rabbit anti-TrkA (1:500; Abcam, Cambridge, UK, 76291), rabbit anti-Lyn Y396ph (1:2000; Abcam, 40660), rabbit anti-Lyn (1:500; CST, 2796s), goat anti-rabbit IgG-HRP (1:2000; Abcam, 97023), goat anti-rabbit IgG-biotin (1:1000; Vector, CA, USA, BA-1000), and Pierce streptavidin poly-HRP (1:3000; Thermo Scientific, MA, USA, 21140).

    Article Title: BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells
    Article Snippet: After blocking, plates were washed three times and the 50 μL undiluted cell culture supernatants were added to 50 μL blocking buffer (non‐fat dry milk) per well. .. Plates were washed 6 times and 1:10 000 streptavidin poly HRP (#21140, Thermo Fisher Scientific) was added and incubated for 1 h at room temperature while shaking (180 rpm).

    other:

    Article Title: Clinical impact of different exosomes’ protein expression in pancreatic ductal carcinoma patients treated with standard first line palliative chemotherapy
    Article Snippet: Streptavidin Poly-HRP (cat. 21140, Thermo Fisher Scientific) is used to amplify the signal.

    Imaging:

    Article Title: Glioblastoma Multiforme Cancer Stem Cells Express Components of the Renin–Angiotensin System
    Article Snippet: ACE tertiary cascade used a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:20,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min. β-actin antibody probing was performed with the iBind™ Flex device (cat# SLF2000, Life Technologies) using primary mouse monoclonal anti-β-actin (1:2000 cat# ab8226, Abcam) and secondary donkey anti-mouse Alexa fluor 488 (1:2000; cat# A21202, Thermo Fisher). .. Clarity Western ECL (cat# 1705061, Bio-Rad) was used as the substrate for visualizing HRP detected protein bands, and the Chemi Doc MP Imaging System (Bio-Rad) and Image Lab 5.0 software (Bio-Rad) were used for both HRP and fluorescent band detection and analysis.

    Article Title: Expression of Components of the Renin-Angiotensin System in Pyogenic Granuloma
    Article Snippet: ACE was probed using a tertiary cascade consisting of a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:4,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min. .. Clarity Western ECL (cat# 1705061, Bio-Rad) was used as the substrate for visualizing HRP detected protein bands and the ChemiDoc MP Imaging System (Bio-Rad) and Image Lab 5.0 software (Bio-Rad) were used for band detection and analysis.

    Sonication:

    Article Title: Lactosylceramide synthases encoded by B4galt5 and 6 genes are pivotal for neuronal generation and myelin formation in mice
    Article Snippet: Western blot analysis Neurospheres cultured on 5 μg/mL laminin-coated glass chambers for differentiation were lysed in NP-40 buffer (150 mM NaCl, 50 mM Tris-Cl, 1% NP-40), containing a phosphatase inhibitor cocktail (Nacalai, Kyoto, Japan), and sonicated with Bioruptor (Diagenode, Liege, Belgium). .. Antibodies and dilutions used in western blotting were as follows: rabbit anti-TrkA Y490ph (1:500; CST, MA, USA, 9141s), rabbit anti-TrkA (1:500; Abcam, Cambridge, UK, 76291), rabbit anti-Lyn Y396ph (1:2000; Abcam, 40660), rabbit anti-Lyn (1:500; CST, 2796s), goat anti-rabbit IgG-HRP (1:2000; Abcam, 97023), goat anti-rabbit IgG-biotin (1:1000; Vector, CA, USA, BA-1000), and Pierce streptavidin poly-HRP (1:3000; Thermo Scientific, MA, USA, 21140).

    Recombinant:

    Article Title: Expression of Components of the Renin-Angiotensin System in Pyogenic Granuloma
    Article Snippet: ACE was probed using a tertiary cascade consisting of a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:4,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min. .. Positive controls were human placenta for PRR and ATIIR1, mouse lung for ACE, and a recombinant ATIIR2 protein (cat# H00000186-P01, Novus Biologicals, Littleton, CO, USA) for ATIIR2.

    Article Title: BAFF augments IgA2 and IL‐10 production by TLR7/8 stimulated total peripheral blood B cells
    Article Snippet: The IgA1 and IgA2 recombinant proteins (16‐16‐090701‐1M and 16‐16‐090701‐2M, Athens Research) were serially diluted in blocking buffer to obtain a standard curve. .. Plates were washed 6 times and 1:10 000 streptavidin poly HRP (#21140, Thermo Fisher Scientific) was added and incubated for 1 h at room temperature while shaking (180 rpm).

    Article Title: ALPPL2 Is a Potential Diagnostic Biomarker for Pancreatic Cancer-Derived Extracellular Vesicles
    Article Snippet: Aptamer-Based Direct and Sandwich ELISA SQ2 aptamer-based direct ELISA was performed with pancreatic cancer cell-derived EVs, concentrated secretome, or human recombinant ALPPL2 protein expressed in HEK293 (OriGene, TP304330) or E. coli (OriGene, SC123000). .. Wells were washed four times with washing buffer, and 2,000-fold diluted streptavidin-poly-HRP conjugate (Pierce) in 0.5% BSA-PBS was added and incubated for 30 min at RT.

    Mutagenesis:

    Article Title: ALPPL2 Is a Potential Diagnostic Biomarker for Pancreatic Cancer-Derived Extracellular Vesicles
    Article Snippet: After blocking, 20 pmol of SQ2 and SQ2 mutant aptamers (St Pharma, South Korea) in binding buffer (PBS with 5 mM MgCl2 , 0.5% BSA, 0.1 mg/mL yeast tRNA, 0.05% Tween-20) were added onto each well and kept for 1 h at RT. .. Wells were washed four times with washing buffer, and 2,000-fold diluted streptavidin-poly-HRP conjugate (Pierce) in 0.5% BSA-PBS was added and incubated for 30 min at RT.

    Labeling:

    Article Title: DNA methyltransferase inhibition overcomes diphthamide pathway deficiencies underlying CD123-targeted treatment resistance
    Article Snippet: Next, slides were incubated in 3% hydrogen peroxide in water for 10 minutes at room temperature, washed 4 times for 5 minutes in PBS with 0.1% Tween-20 (PBS-T), incubated with protein block (Leica, RE7102) for 30 minutes at room temperature, and then washed 4 times for 5 minutes in PBS-T. For enzymatic labeling, cells were incubated in a buffer containing 20 mM Tris-Cl pH 7.4, 50 mM DTT, 50 μM biotinylated NAD+ (Trevigen, 4670), with or without 20 ng/μL tagraxofusp at 37°C for 1 hour in a humid box. .. After incubation, slides were washed 4 times for 5 minutes each in PBS-T, and then incubated with streptavidin-poly HRP (Thermo Fisher Scientific, 21140) in PBS containing 1% BSA for 30 minutes at room temperature in a humid box.

    Purification:

    Article Title: Utilizing Biotinylated Proteins Expressed in Yeast to Visualize DNA–Protein Interactions at the Single-Molecule Level
    Article Snippet: Boiled whole-protein extracts or purified protein samples were incubated with unboiled streptavidin for 5 min at room temperature and then loaded onto a sodium dodecyl sulfate-polyacrylamide gel. .. The antibodies used in this study were CBP tag antibody (A01798, Genscript), Flag tag antibody (F3165, Sigma-Aldrich), and streptavidin poly-HRP (21140, Thermo Fisher Scientific).

    Protein Extraction:

    Article Title: Lactosylceramide synthases encoded by B4galt5 and 6 genes are pivotal for neuronal generation and myelin formation in mice
    Article Snippet: Antibodies and dilutions used in western blotting were as follows: rabbit anti-TrkA Y490ph (1:500; CST, MA, USA, 9141s), rabbit anti-TrkA (1:500; Abcam, Cambridge, UK, 76291), rabbit anti-Lyn Y396ph (1:2000; Abcam, 40660), rabbit anti-Lyn (1:500; CST, 2796s), goat anti-rabbit IgG-HRP (1:2000; Abcam, 97023), goat anti-rabbit IgG-biotin (1:1000; Vector, CA, USA, BA-1000), and Pierce streptavidin poly-HRP (1:3000; Thermo Scientific, MA, USA, 21140). .. Western blotting for brain homogenates was carried out, as described previously [ ], using total brain proteins, extracted using total protein extraction kit (P501S; 101 Bio, AC, USA).

    Direct ELISA:

    Article Title: ALPPL2 Is a Potential Diagnostic Biomarker for Pancreatic Cancer-Derived Extracellular Vesicles
    Article Snippet: Aptamer-Based Direct and Sandwich ELISA SQ2 aptamer-based direct ELISA was performed with pancreatic cancer cell-derived EVs, concentrated secretome, or human recombinant ALPPL2 protein expressed in HEK293 (OriGene, TP304330) or E. coli (OriGene, SC123000). .. Wells were washed four times with washing buffer, and 2,000-fold diluted streptavidin-poly-HRP conjugate (Pierce) in 0.5% BSA-PBS was added and incubated for 30 min at RT.

    Electrophoretic Mobility Shift Assay:

    Article Title: Utilizing Biotinylated Proteins Expressed in Yeast to Visualize DNA–Protein Interactions at the Single-Molecule Level
    Article Snippet: Biotinylation detection and efficiency evaluation The biotinylation of target proteins was detected using streptavidin-horseradish peroxidase (HRP); the extent of biotinylation was evaluated using a streptavidin gel shift assay as described by van Werven and Timmers ( ). .. The antibodies used in this study were CBP tag antibody (A01798, Genscript), Flag tag antibody (F3165, Sigma-Aldrich), and streptavidin poly-HRP (21140, Thermo Fisher Scientific).

    Polyacrylamide Gel Electrophoresis:

    Article Title: Expression of Components of the Renin-Angiotensin System in Pyogenic Granuloma
    Article Snippet: The protein extracts were resolved by 4–12% one-dimensional polyacrylamide gel electrophoresis then transferred to polyvinylidene difluoride membranes. .. ACE was probed using a tertiary cascade consisting of a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:4,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min.

    SDS Page:

    Article Title: Lactosylceramide synthases encoded by B4galt5 and 6 genes are pivotal for neuronal generation and myelin formation in mice
    Article Snippet: The samples were applied on 5–12% SuperSep SDS-PAGE gel (Wako, Osaka, Japan) and transferred to PVDF membrane (Merck Millipore, Darmstadt, Germany). .. Antibodies and dilutions used in western blotting were as follows: rabbit anti-TrkA Y490ph (1:500; CST, MA, USA, 9141s), rabbit anti-TrkA (1:500; Abcam, Cambridge, UK, 76291), rabbit anti-Lyn Y396ph (1:2000; Abcam, 40660), rabbit anti-Lyn (1:500; CST, 2796s), goat anti-rabbit IgG-HRP (1:2000; Abcam, 97023), goat anti-rabbit IgG-biotin (1:1000; Vector, CA, USA, BA-1000), and Pierce streptavidin poly-HRP (1:3000; Thermo Scientific, MA, USA, 21140).

    Plasmid Preparation:

    Article Title: DNA methyltransferase inhibition overcomes diphthamide pathway deficiencies underlying CD123-targeted treatment resistance
    Article Snippet: After incubation, slides were washed 4 times for 5 minutes each in PBS-T, and then incubated with streptavidin-poly HRP (Thermo Fisher Scientific, 21140) in PBS containing 1% BSA for 30 minutes at room temperature in a humid box. .. After incubation, slides were washed 4 times for 5 minutes each in PBS-T, and then incubated with streptavidin-poly HRP (Thermo Fisher Scientific, 21140) in PBS containing 1% BSA for 30 minutes at room temperature in a humid box.

    Article Title: Lactosylceramide synthases encoded by B4galt5 and 6 genes are pivotal for neuronal generation and myelin formation in mice
    Article Snippet: .. Antibodies and dilutions used in western blotting were as follows: rabbit anti-TrkA Y490ph (1:500; CST, MA, USA, 9141s), rabbit anti-TrkA (1:500; Abcam, Cambridge, UK, 76291), rabbit anti-Lyn Y396ph (1:2000; Abcam, 40660), rabbit anti-Lyn (1:500; CST, 2796s), goat anti-rabbit IgG-HRP (1:2000; Abcam, 97023), goat anti-rabbit IgG-biotin (1:1000; Vector, CA, USA, BA-1000), and Pierce streptavidin poly-HRP (1:3000; Thermo Scientific, MA, USA, 21140). .. Western blotting for brain homogenates was carried out, as described previously [ ], using total brain proteins, extracted using total protein extraction kit (P501S; 101 Bio, AC, USA).

    Article Title: An ABCA1-independent pathway for recycling a poorly lipidated 8.1 nm apolipoprotein E particle from glia
    Article Snippet: After washing, 100 µl per well of biotinylated anti-goat IgG (1:160,000; Vector) in DB was added and incubated for 90 min at 37°C. .. Streptavidin-poly-HRP antibody (1:4000; Pierce) was then added and incubated at RT for 90 min. Color was developed by adding 100 μl/well Sigma ELISA TMB and stopped immediately by adding 100 μl of 1M HCL.

    Software:

    Article Title: Glioblastoma Multiforme Cancer Stem Cells Express Components of the Renin–Angiotensin System
    Article Snippet: ACE tertiary cascade used a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:20,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min. β-actin antibody probing was performed with the iBind™ Flex device (cat# SLF2000, Life Technologies) using primary mouse monoclonal anti-β-actin (1:2000 cat# ab8226, Abcam) and secondary donkey anti-mouse Alexa fluor 488 (1:2000; cat# A21202, Thermo Fisher). .. Clarity Western ECL (cat# 1705061, Bio-Rad) was used as the substrate for visualizing HRP detected protein bands, and the Chemi Doc MP Imaging System (Bio-Rad) and Image Lab 5.0 software (Bio-Rad) were used for both HRP and fluorescent band detection and analysis.

    Article Title: Expression of Components of the Renin-Angiotensin System in Pyogenic Granuloma
    Article Snippet: ACE was probed using a tertiary cascade consisting of a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:4,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min. .. Clarity Western ECL (cat# 1705061, Bio-Rad) was used as the substrate for visualizing HRP detected protein bands and the ChemiDoc MP Imaging System (Bio-Rad) and Image Lab 5.0 software (Bio-Rad) were used for band detection and analysis.

    Binding Assay:

    Article Title: ALPPL2 Is a Potential Diagnostic Biomarker for Pancreatic Cancer-Derived Extracellular Vesicles
    Article Snippet: After blocking, 20 pmol of SQ2 and SQ2 mutant aptamers (St Pharma, South Korea) in binding buffer (PBS with 5 mM MgCl2 , 0.5% BSA, 0.1 mg/mL yeast tRNA, 0.05% Tween-20) were added onto each well and kept for 1 h at RT. .. Wells were washed four times with washing buffer, and 2,000-fold diluted streptavidin-poly-HRP conjugate (Pierce) in 0.5% BSA-PBS was added and incubated for 30 min at RT.

    FLAG-tag:

    Article Title: Utilizing Biotinylated Proteins Expressed in Yeast to Visualize DNA–Protein Interactions at the Single-Molecule Level
    Article Snippet: .. The antibodies used in this study were CBP tag antibody (A01798, Genscript), Flag tag antibody (F3165, Sigma-Aldrich), and streptavidin poly-HRP (21140, Thermo Fisher Scientific). .. For purified proteins, the gel was stained using Coomassie Brilliant Blue, and the intensity of the bands was quantified using Image J.

    Marker:

    Article Title: Glioblastoma Multiforme Cancer Stem Cells Express Components of the Renin–Angiotensin System
    Article Snippet: Primary antibody probing for each RAS marker was overnight in TBST at 4°C with the following primary antibodies at the given concentrations: PRR (ATP6IP2, 1:500, cat# ab40790, Abcam, Cambridge, UK), ATIIR1 (AT2R1, 1:500; cat# sc-1173, Santa Cruz, CA, USA), ATIIR2 (1:5000; cat# ab92445, Abcam), and ACE (1:200; cat# sc-12184, Santa Cruz). .. ACE tertiary cascade used a rabbit anti-goat Superclonal™ biotin conjugated secondary antibody (1:20,000; cat# A27013, Thermo Fisher) followed by a Pierce™ Streptavidin Poly HRP (1:5000, cat# 21140, Thermo Fisher) at 4°C for 10 min. β-actin antibody probing was performed with the iBind™ Flex device (cat# SLF2000, Life Technologies) using primary mouse monoclonal anti-β-actin (1:2000 cat# ab8226, Abcam) and secondary donkey anti-mouse Alexa fluor 488 (1:2000; cat# A21202, Thermo Fisher).

    Staining:

    Article Title: Utilizing Biotinylated Proteins Expressed in Yeast to Visualize DNA–Protein Interactions at the Single-Molecule Level
    Article Snippet: The antibodies used in this study were CBP tag antibody (A01798, Genscript), Flag tag antibody (F3165, Sigma-Aldrich), and streptavidin poly-HRP (21140, Thermo Fisher Scientific). .. For purified proteins, the gel was stained using Coomassie Brilliant Blue, and the intensity of the bands was quantified using Image J.

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    Thermo Fisher streptavidin poly hrp
    Schematic of an amplification strategy using <t>streptavidin</t> <t>poly-HRP.</t> The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.
    Streptavidin Poly Hrp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.

    Journal: Lab on a Chip

    Article Title: Fabrication of immunosensor microwell arrays from gold compact discs for detection of cancer biomarker proteins †

    doi: 10.1039/c1lc20833k

    Figure Lengend Snippet: Schematic of an amplification strategy using streptavidin poly-HRP. The streptavidin poly-HRP is attached to the biotinylated anti-human IL-6 detection antibody.

    Article Snippet: In this work, we chose an alternative, rapid amplification strategy that substitutes streptavidin poly-HRP for the more commonly used streptavidin HRP to improve sensitivity and limit of detection.

    Techniques: Amplification

    Lectin array analysis of exposed glycans on rFVIII products. rFVIII products were adsorbed on microtitre plates at 1 μg/mL and assessed using a panel of biotinylated lectins. BSA was adsorbed as a control. Binding was detected using a streptavidin poly-HRP and read at 492 nm. (A) Heat plot demonstrating the median-centred binding values of lectins with varying carbohydrate specificites to different rFVIII products. Values shown are representative of the mean of at least 3 independent experiments on a single lot of each rFVIII product. Statistical summary available in Online Supplementary Table S1 . (B) Lectin binding analysis of different production lots of full-length rFVIII from CHO or BHK cells. Data are representative of at least 3 independent experiments. BHK: baby hamster kidney cells; CHO: Chinese hamster ovary cells: rFVIII: recombinant factor VIII; BSA: bovine serum albumin; BDD: B-domain deleted.

    Journal: Haematologica

    Article Title: N-linked glycosylation modulates the immunogenicity of recombinant human factor VIII in hemophilia A mice

    doi: 10.3324/haematol.2018.188219

    Figure Lengend Snippet: Lectin array analysis of exposed glycans on rFVIII products. rFVIII products were adsorbed on microtitre plates at 1 μg/mL and assessed using a panel of biotinylated lectins. BSA was adsorbed as a control. Binding was detected using a streptavidin poly-HRP and read at 492 nm. (A) Heat plot demonstrating the median-centred binding values of lectins with varying carbohydrate specificites to different rFVIII products. Values shown are representative of the mean of at least 3 independent experiments on a single lot of each rFVIII product. Statistical summary available in Online Supplementary Table S1 . (B) Lectin binding analysis of different production lots of full-length rFVIII from CHO or BHK cells. Data are representative of at least 3 independent experiments. BHK: baby hamster kidney cells; CHO: Chinese hamster ovary cells: rFVIII: recombinant factor VIII; BSA: bovine serum albumin; BDD: B-domain deleted.

    Article Snippet: Detection was facilitated using streptavidin-poly-HRP (ThermoFisher Scientific, Waltham, MA, USA) and developed for 5 min. Statistical analysis was performed using the Student t test.

    Techniques: Binding Assay, Recombinant