dynabeads m 280 streptavidin  (Thermo Fisher)


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    Structured Review

    Thermo Fisher dynabeads m 280 streptavidin
    Dynabeads M 280 Streptavidin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dynabeads m 280 streptavidin/product/Thermo Fisher
    Average 99 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    dynabeads m 280 streptavidin - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Centrifugation:

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C. .. The next day, a beads-probes-RNAs mixture was washed and incubated with 200 μl lysis buffer and proteinase K to reverse the formaldehyde crosslinking.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C. .. The next day, a beads-probes-RNAs mixture was washed and incubated with 200 μl lysis buffer and proteinase K to reverse the formaldehyde crosslinking.

    Amplification:

    Article Title: Full-length RNA profiling reveals pervasive bidirectional transcription terminators in bacteria
    Article Snippet: Biotin-labeled DNA strands were bound to the Dynabeads M-280 Streptavidin (Invitrogen, 11205D) and cleaned four times with Washing Buffer (5 mM Tris-HCl pH 7.5, 1 M NaCl, 0.5 mM EDTA) and twice with TE buffer. .. The DNA library was then amplified for 13 (total RNA SEnd-seq) to 17 cycles (primary RNA SEnd-seq) following the manufacturer’s protocol.

    Polymerase Chain Reaction:

    Article Title: LINC02273 drives breast cancer metastasis by epigenetically increasing AGR2 transcription
    Article Snippet: RNA pull-down assay PCR products were verified by gel electrophoresis and purified by phenol: chloroform extraction. .. For each sample, 5 μg RNA was mixed with 1 × 107 cell extract and incubated at 4 °C for 1 h, followed by incubating with Dynabeads M-280 Streptavidin (Invitrogen) at 4 °C overnight.

    Quantitative RT-PCR:

    Article Title: Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p, et al. Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p
    Article Snippet: Then, 30 μL streptavidin‐conjugated magnetic beads (11205D, Invitrogen) were added into the cell lysate and incubated at 4°C for 1 hour. .. The retrieved RNA was detected by qRT‐PCR assay as described above.

    Incubation:

    Article Title: Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p, et al. Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p
    Article Snippet: .. Then, 30 μL streptavidin‐conjugated magnetic beads (11205D, Invitrogen) were added into the cell lysate and incubated at 4°C for 1 hour. .. The retrieved RNA was detected by qRT‐PCR assay as described above.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C. .. The next day, a beads-probes-RNAs mixture was washed and incubated with 200 μl lysis buffer and proteinase K to reverse the formaldehyde crosslinking.

    Article Title: lncRNA MALAT1 binds chromatin remodeling subunit BRG1 to epigenetically promote inflammation-related hepatocellular carcinoma progression
    Article Snippet: .. The second day, 50 µl washed Streptavidin agarose beads (11206D, Invitrogen) were added to each binding reaction and incubated at 4 °C for another one hour. ..

    Article Title: LINC02273 drives breast cancer metastasis by epigenetically increasing AGR2 transcription
    Article Snippet: .. For each sample, 5 μg RNA was mixed with 1 × 107 cell extract and incubated at 4 °C for 1 h, followed by incubating with Dynabeads M-280 Streptavidin (Invitrogen) at 4 °C overnight. ..

    Article Title: The long noncoding RNA LOC105374325 causes podocyte injury in individuals with focal segmental glomerulosclerosis
    Article Snippet: .. The biotinylated DNA probe complementary to LOC105374325 (100 pmol) was incubated with Dynabeads M-280 Streptavidin (11205D, Invitrogen) at room temperature for 10 min to generate probe-coated beads according to the manufacturer's protocol. .. Then, tissue or cell lysates were incubated with the probe-coated beads, and the RNA complexes bound to these beads were extracted for RT-PCR analysis ( , ).

    Article Title: The Orphan Immune Receptor LILRB3 Modulates Fc Receptor-Mediated Functions of Neutrophils.
    Article Snippet: .. Neutrophils are critical to the generation of effective immune responses and for killing invading microbes. ..

    Article Title: Targeting the tumor vasculature with engineered cystine-knot miniproteins
    Article Snippet: For each screening round 2 × 50 μL Dynabeads® M-280 Streptavidin (Life Technologies) were transferred to a 2 mL tube each and washed with 1 mL TBS-T buffer (50 mM Tris, 150 mM NaCl, 0.1% Tween-20, pH 7.4). .. 100 µg biotinylated FN-B in 200 μL TBS (50 mM Tris, 150 mM NaCl, pH 7.4) was added to the first tube, 200 μL TBS without target protein to the second tube (used for negative selection of phages) and beads were incubated on a rolling mixer for 20 min at 30 rpm.

    Article Title: A polyomavirus peptide binds to the capsid VP1 pore and has potent antiviral activity against BK and JC polyomaviruses
    Article Snippet: Peptide-virus co-purification assay 54 nM peptide was incubated with 75 µg/mL (370 nM) recombinant full-length VP1 pentamers, or molar equivalent of VP1 pentamers of BK VLP or purified infectious BKV (as assessed by SDS-PAGE and comparing VP1 abundance by Coomassie stain) in 336 μl PBS buffer containing 1% DMSO and 0.01% Tween-20 for 1 hr at room temperature. .. 300 μl of mix was added to a fresh tube, and 20 μl of Dynabeads M-280 Streptavidin (Invitrogen) were added.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C. .. The next day, a beads-probes-RNAs mixture was washed and incubated with 200 μl lysis buffer and proteinase K to reverse the formaldehyde crosslinking.

    Mass Spectrometry:

    Article Title: lncRNA MALAT1 binds chromatin remodeling subunit BRG1 to epigenetically promote inflammation-related hepatocellular carcinoma progression
    Article Snippet: The second day, 50 µl washed Streptavidin agarose beads (11206D, Invitrogen) were added to each binding reaction and incubated at 4 °C for another one hour. .. Then the denatured proteins were detected by western blot or resolved in gradient gel electrophoresis followed by mass spectrometry (MS) identification.

    BIA-KA:

    Article Title: Cell-specific non-canonical amino acid labelling identifies changes in the de novo proteome during memory formation
    Article Snippet: The samples were snap-frozen and then extracted in radioimmunoprecipitation assay (RIPA) buffer (Cell Signalling, 9806) as previously described , with protein concentrations being determined using the bicinchoninic acid (BCA) assay (ThermoFisher, 23225). .. 40 μg of streptavidin-coated Dynabeads (ThermoFisher, 11205D) were then used to purify biotinylated proteins, with beads being washed multiple times with IP wash buffer (0.1% SDS and 0.05% Tween in Tris-buffered saline (TBS)).

    Modification:

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: According to ), with modification, biotin-labeled miRNA-1224 probe (Bio-1224, 5′-CT CCACCTCCCCAGTCCTCAC-Bio-3′) was used to perform the RRIP experiment assay. .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: According to ), with modification, biotin-labeled miRNA-1224 probe (Bio-1224, 5′-CT CCACCTCCCCAGTCCTCAC-Bio-3′) was used to perform the RRIP experiment assay. .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C.

    Western Blot:

    Article Title: lncRNA MALAT1 binds chromatin remodeling subunit BRG1 to epigenetically promote inflammation-related hepatocellular carcinoma progression
    Article Snippet: The second day, 50 µl washed Streptavidin agarose beads (11206D, Invitrogen) were added to each binding reaction and incubated at 4 °C for another one hour. .. Then the denatured proteins were detected by western blot or resolved in gradient gel electrophoresis followed by mass spectrometry (MS) identification.

    Article Title: Cell-specific non-canonical amino acid labelling identifies changes in the de novo proteome during memory formation
    Article Snippet: Briefly, for samples to be analysed via western blot following BONCAT purification (BONCAT-WB), 100 μg of protein was used. .. 40 μg of streptavidin-coated Dynabeads (ThermoFisher, 11205D) were then used to purify biotinylated proteins, with beads being washed multiple times with IP wash buffer (0.1% SDS and 0.05% Tween in Tris-buffered saline (TBS)).

    Flow Cytometry:

    Article Title: The Orphan Immune Receptor LILRB3 Modulates Fc Receptor-Mediated Functions of Neutrophils.
    Article Snippet: Neutrophils are critical to the generation of effective immune responses and for killing invading microbes. .. Neutrophils are critical to the generation of effective immune responses and for killing invading microbes.

    Protease Inhibitor:

    Article Title: Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p, et al. Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p
    Article Snippet: Cellular protein was extracted using lysis buffer [100 mmol/L KCl, 5 mmol/L MgCl2 , 10 mmol/L HEPES (pH 7.0), 0.5% NP‐40 supplemented with fresh 200 U RNase inhibitor (Yeasen Biotech Co Ltd), 1 mmol/L DTT, 20 mmol/L EDTA, EDTA‐free protease inhibitor cocktail (Roche) and PMSF] and incubated with 3 μg biotin‐coupled probes at 4°C for 2 hours. .. Then, 30 μL streptavidin‐conjugated magnetic beads (11205D, Invitrogen) were added into the cell lysate and incubated at 4°C for 1 hour.

    Article Title: Phage antibody library screening for the selection of novel high-affinity human single-chain variable fragment against gastrin receptor: an in silico and in vitro study
    Article Snippet: The secondary goat anti-mouse IgG conjugated HRP and Dynabeads M-280 Streptavidin were obtained from Invitrogen Life Technology (Karlsruhe, Germany). .. ULTRA tablets and protease inhibitor cocktail were obtained from Roche (Basel, Switzerland).

    Sequencing:

    Article Title: Full-length RNA profiling reveals pervasive bidirectional transcription terminators in bacteria
    Article Snippet: The DNA ends were prepared and ligated to the Illumina sequencing adaptor with the NEBNext Ultra II DNA Library Prep Kit (New England BioLabs, E7645). .. Biotin-labeled DNA strands were bound to the Dynabeads M-280 Streptavidin (Invitrogen, 11205D) and cleaned four times with Washing Buffer (5 mM Tris-HCl pH 7.5, 1 M NaCl, 0.5 mM EDTA) and twice with TE buffer.

    Sonication:

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: Spinal cord was harvested 24 h after intrathecal injection of Bio-1224 (5 μl, 20 μ m ) and fixed by 2.5% formaldehyde for 10 min, lysed, and sonicated. .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: Spinal cord was harvested 24 h after intrathecal injection of Bio-1224 (5 μl, 20 μ m ) and fixed by 2.5% formaldehyde for 10 min, lysed, and sonicated. .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C.

    Injection:

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: Spinal cord was harvested 24 h after intrathecal injection of Bio-1224 (5 μl, 20 μ m ) and fixed by 2.5% formaldehyde for 10 min, lysed, and sonicated. .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: Spinal cord was harvested 24 h after intrathecal injection of Bio-1224 (5 μl, 20 μ m ) and fixed by 2.5% formaldehyde for 10 min, lysed, and sonicated. .. After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C.

    Copurification:

    Article Title: A polyomavirus peptide binds to the capsid VP1 pore and has potent antiviral activity against BK and JC polyomaviruses
    Article Snippet: Paragraph title: Peptide-virus co-purification assay ... 300 μl of mix was added to a fresh tube, and 20 μl of Dynabeads M-280 Streptavidin (Invitrogen) were added.

    Nucleic Acid Electrophoresis:

    Article Title: lncRNA MALAT1 binds chromatin remodeling subunit BRG1 to epigenetically promote inflammation-related hepatocellular carcinoma progression
    Article Snippet: The second day, 50 µl washed Streptavidin agarose beads (11206D, Invitrogen) were added to each binding reaction and incubated at 4 °C for another one hour. .. Then the denatured proteins were detected by western blot or resolved in gradient gel electrophoresis followed by mass spectrometry (MS) identification.

    Article Title: LINC02273 drives breast cancer metastasis by epigenetically increasing AGR2 transcription
    Article Snippet: RNA pull-down assay PCR products were verified by gel electrophoresis and purified by phenol: chloroform extraction. .. For each sample, 5 μg RNA was mixed with 1 × 107 cell extract and incubated at 4 °C for 1 h, followed by incubating with Dynabeads M-280 Streptavidin (Invitrogen) at 4 °C overnight.

    In Vivo:

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: Paragraph title: RNA-RNA in vivo precipitation (RRIP). ... After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C.

    Pull Down Assay:

    Article Title: LINC02273 drives breast cancer metastasis by epigenetically increasing AGR2 transcription
    Article Snippet: Paragraph title: RNA pull-down assay ... For each sample, 5 μg RNA was mixed with 1 × 107 cell extract and incubated at 4 °C for 1 h, followed by incubating with Dynabeads M-280 Streptavidin (Invitrogen) at 4 °C overnight.

    Magnetic Beads:

    Article Title: Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p, et al. Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p
    Article Snippet: .. Then, 30 μL streptavidin‐conjugated magnetic beads (11205D, Invitrogen) were added into the cell lysate and incubated at 4°C for 1 hour. .. The retrieved RNA was detected by qRT‐PCR assay as described above.

    Article Title: Targeting the tumor vasculature with engineered cystine-knot miniproteins
    Article Snippet: In total, three screening rounds based on streptavidin-coated (SA) magnetic beads were performed with each library. .. For each screening round 2 × 50 μL Dynabeads® M-280 Streptavidin (Life Technologies) were transferred to a 2 mL tube each and washed with 1 mL TBS-T buffer (50 mM Tris, 150 mM NaCl, 0.1% Tween-20, pH 7.4).

    Size-exclusion Chromatography:

    Article Title: Full-length RNA profiling reveals pervasive bidirectional transcription terminators in bacteria
    Article Snippet: Circularized cDNA was fragmented by acoustic shearing in microTUBE (Covaris, 520045) with Covaris S220 Focused-ultrasonicator under the condition of Peak145 for 90 sec. After ethanol precipitation, the ssDNA was converted to dsDNA by the Second Strand cDNA Synthesis Kit (New England BioLabs, E6114) at 16 °C for 2 hr. .. Biotin-labeled DNA strands were bound to the Dynabeads M-280 Streptavidin (Invitrogen, 11205D) and cleaned four times with Washing Buffer (5 mM Tris-HCl pH 7.5, 1 M NaCl, 0.5 mM EDTA) and twice with TE buffer.

    Labeling:

    Article Title: lncRNA MALAT1 binds chromatin remodeling subunit BRG1 to epigenetically promote inflammation-related hepatocellular carcinoma progression
    Article Snippet: Biotin RNA Labeling Mix (11685597910, Roche) and T7 RNA polymerase (10881767001, Roche) were applied to synthesize biotin-labeled RNAs, and then purified with RNeasy Mini Kit (74104, Qiagen) after treated with RNase-free DNase I (M610A, Promega). .. The second day, 50 µl washed Streptavidin agarose beads (11206D, Invitrogen) were added to each binding reaction and incubated at 4 °C for another one hour.

    Article Title: The Orphan Immune Receptor LILRB3 Modulates Fc Receptor-Mediated Functions of Neutrophils.
    Article Snippet: .. Neutrophils are critical to the generation of effective immune responses and for killing invading microbes. ..

    Purification:

    Article Title: lncRNA MALAT1 binds chromatin remodeling subunit BRG1 to epigenetically promote inflammation-related hepatocellular carcinoma progression
    Article Snippet: Biotin RNA Labeling Mix (11685597910, Roche) and T7 RNA polymerase (10881767001, Roche) were applied to synthesize biotin-labeled RNAs, and then purified with RNeasy Mini Kit (74104, Qiagen) after treated with RNase-free DNase I (M610A, Promega). .. The second day, 50 µl washed Streptavidin agarose beads (11206D, Invitrogen) were added to each binding reaction and incubated at 4 °C for another one hour.

    Article Title: Cell-specific non-canonical amino acid labelling identifies changes in the de novo proteome during memory formation
    Article Snippet: Paragraph title: BONCAT purification ... 40 μg of streptavidin-coated Dynabeads (ThermoFisher, 11205D) were then used to purify biotinylated proteins, with beads being washed multiple times with IP wash buffer (0.1% SDS and 0.05% Tween in Tris-buffered saline (TBS)).

    Article Title: LINC02273 drives breast cancer metastasis by epigenetically increasing AGR2 transcription
    Article Snippet: Biotin-labelled RNAs was in vitro transcribed with HiScribe™ T7 Quick High Yield RNA Synthesis Kit (NEB) with Biotin-16-UTP (Roche), treated with RNase-free DNase I on column during RNA purification with RNA Clean & Concentrator-25 (Zymo Research). .. For each sample, 5 μg RNA was mixed with 1 × 107 cell extract and incubated at 4 °C for 1 h, followed by incubating with Dynabeads M-280 Streptavidin (Invitrogen) at 4 °C overnight.

    Article Title: A polyomavirus peptide binds to the capsid VP1 pore and has potent antiviral activity against BK and JC polyomaviruses
    Article Snippet: Peptide-virus co-purification assay 54 nM peptide was incubated with 75 µg/mL (370 nM) recombinant full-length VP1 pentamers, or molar equivalent of VP1 pentamers of BK VLP or purified infectious BKV (as assessed by SDS-PAGE and comparing VP1 abundance by Coomassie stain) in 336 μl PBS buffer containing 1% DMSO and 0.01% Tween-20 for 1 hr at room temperature. .. 300 μl of mix was added to a fresh tube, and 20 μl of Dynabeads M-280 Streptavidin (Invitrogen) were added.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: The long noncoding RNA LOC105374325 causes podocyte injury in individuals with focal segmental glomerulosclerosis
    Article Snippet: The biotinylated DNA probe complementary to LOC105374325 (100 pmol) was incubated with Dynabeads M-280 Streptavidin (11205D, Invitrogen) at room temperature for 10 min to generate probe-coated beads according to the manufacturer's protocol. .. Then, tissue or cell lysates were incubated with the probe-coated beads, and the RNA complexes bound to these beads were extracted for RT-PCR analysis ( , ).

    Protein Extraction:

    Article Title: lncRNA MALAT1 binds chromatin remodeling subunit BRG1 to epigenetically promote inflammation-related hepatocellular carcinoma progression
    Article Snippet: Folded RNA was then mixed with QGY-7701/QGY-7703 nuclear protein extraction (containing 1 mg proteins) in IP buffer and then incubated at 4 °C for overnight. .. The second day, 50 µl washed Streptavidin agarose beads (11206D, Invitrogen) were added to each binding reaction and incubated at 4 °C for another one hour.

    Selection:

    Article Title: Targeting the tumor vasculature with engineered cystine-knot miniproteins
    Article Snippet: Paragraph title: Selection of EDB-specific ligands via phage display ... For each screening round 2 × 50 μL Dynabeads® M-280 Streptavidin (Life Technologies) were transferred to a 2 mL tube each and washed with 1 mL TBS-T buffer (50 mM Tris, 150 mM NaCl, 0.1% Tween-20, pH 7.4).

    Construct:

    Article Title: Targeting the tumor vasculature with engineered cystine-knot miniproteins
    Article Snippet: Selection of EDB-specific ligands via phage display Two different combinatorial libraries, which are based on oMCoTI-II were constructed using an M13 phagemid system. .. For each screening round 2 × 50 μL Dynabeads® M-280 Streptavidin (Life Technologies) were transferred to a 2 mL tube each and washed with 1 mL TBS-T buffer (50 mM Tris, 150 mM NaCl, 0.1% Tween-20, pH 7.4).

    Staining:

    Article Title: A polyomavirus peptide binds to the capsid VP1 pore and has potent antiviral activity against BK and JC polyomaviruses
    Article Snippet: Peptide-virus co-purification assay 54 nM peptide was incubated with 75 µg/mL (370 nM) recombinant full-length VP1 pentamers, or molar equivalent of VP1 pentamers of BK VLP or purified infectious BKV (as assessed by SDS-PAGE and comparing VP1 abundance by Coomassie stain) in 336 μl PBS buffer containing 1% DMSO and 0.01% Tween-20 for 1 hr at room temperature. .. 300 μl of mix was added to a fresh tube, and 20 μl of Dynabeads M-280 Streptavidin (Invitrogen) were added.

    SDS Page:

    Article Title: A polyomavirus peptide binds to the capsid VP1 pore and has potent antiviral activity against BK and JC polyomaviruses
    Article Snippet: Peptide-virus co-purification assay 54 nM peptide was incubated with 75 µg/mL (370 nM) recombinant full-length VP1 pentamers, or molar equivalent of VP1 pentamers of BK VLP or purified infectious BKV (as assessed by SDS-PAGE and comparing VP1 abundance by Coomassie stain) in 336 μl PBS buffer containing 1% DMSO and 0.01% Tween-20 for 1 hr at room temperature. .. 300 μl of mix was added to a fresh tube, and 20 μl of Dynabeads M-280 Streptavidin (Invitrogen) were added.

    RNA Extraction:

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C. .. Finally, the mixture was added with TRizol for RNA extraction and detection.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C. .. Finally, the mixture was added with TRizol for RNA extraction and detection.

    Binding Assay:

    Article Title: lncRNA MALAT1 binds chromatin remodeling subunit BRG1 to epigenetically promote inflammation-related hepatocellular carcinoma progression
    Article Snippet: .. The second day, 50 µl washed Streptavidin agarose beads (11206D, Invitrogen) were added to each binding reaction and incubated at 4 °C for another one hour. ..

    In Vitro:

    Article Title: LINC02273 drives breast cancer metastasis by epigenetically increasing AGR2 transcription
    Article Snippet: Biotin-labelled RNAs was in vitro transcribed with HiScribe™ T7 Quick High Yield RNA Synthesis Kit (NEB) with Biotin-16-UTP (Roche), treated with RNase-free DNase I on column during RNA purification with RNA Clean & Concentrator-25 (Zymo Research). .. For each sample, 5 μg RNA was mixed with 1 × 107 cell extract and incubated at 4 °C for 1 h, followed by incubating with Dynabeads M-280 Streptavidin (Invitrogen) at 4 °C overnight.

    Radio Immunoprecipitation:

    Article Title: Cell-specific non-canonical amino acid labelling identifies changes in the de novo proteome during memory formation
    Article Snippet: The samples were snap-frozen and then extracted in radioimmunoprecipitation assay (RIPA) buffer (Cell Signalling, 9806) as previously described , with protein concentrations being determined using the bicinchoninic acid (BCA) assay (ThermoFisher, 23225). .. 40 μg of streptavidin-coated Dynabeads (ThermoFisher, 11205D) were then used to purify biotinylated proteins, with beads being washed multiple times with IP wash buffer (0.1% SDS and 0.05% Tween in Tris-buffered saline (TBS)).

    Ethanol Precipitation:

    Article Title: Full-length RNA profiling reveals pervasive bidirectional transcription terminators in bacteria
    Article Snippet: Circularized cDNA was fragmented by acoustic shearing in microTUBE (Covaris, 520045) with Covaris S220 Focused-ultrasonicator under the condition of Peak145 for 90 sec. After ethanol precipitation, the ssDNA was converted to dsDNA by the Second Strand cDNA Synthesis Kit (New England BioLabs, E6114) at 16 °C for 2 hr. .. Biotin-labeled DNA strands were bound to the Dynabeads M-280 Streptavidin (Invitrogen, 11205D) and cleaned four times with Washing Buffer (5 mM Tris-HCl pH 7.5, 1 M NaCl, 0.5 mM EDTA) and twice with TE buffer.

    Concentration Assay:

    Article Title: Full-length RNA profiling reveals pervasive bidirectional transcription terminators in bacteria
    Article Snippet: Biotin-labeled DNA strands were bound to the Dynabeads M-280 Streptavidin (Invitrogen, 11205D) and cleaned four times with Washing Buffer (5 mM Tris-HCl pH 7.5, 1 M NaCl, 0.5 mM EDTA) and twice with TE buffer. .. The final library was cleaned twice with 1× vol (50 μl) of AMPure beads, and its concentration and size distribution were determined with Agilent 2200 TapeStation (Agilent, 5067–5576).

    Lysis:

    Article Title: Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p, et al. Circular RNA circFOXO3 promotes prostate cancer progression through sponging miR‐29a‐3p
    Article Snippet: Cellular protein was extracted using lysis buffer [100 mmol/L KCl, 5 mmol/L MgCl2 , 10 mmol/L HEPES (pH 7.0), 0.5% NP‐40 supplemented with fresh 200 U RNase inhibitor (Yeasen Biotech Co Ltd), 1 mmol/L DTT, 20 mmol/L EDTA, EDTA‐free protease inhibitor cocktail (Roche) and PMSF] and incubated with 3 μg biotin‐coupled probes at 4°C for 2 hours. .. Then, 30 μL streptavidin‐conjugated magnetic beads (11205D, Invitrogen) were added into the cell lysate and incubated at 4°C for 1 hour.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C. .. The next day, a beads-probes-RNAs mixture was washed and incubated with 200 μl lysis buffer and proteinase K to reverse the formaldehyde crosslinking.

    Article Title: MicroRNA-1224 Splicing CircularRNA-Filip1l in an Ago2-Dependent Manner Regulates Chronic Inflammatory Pain via Targeting Ubr5
    Article Snippet: After centrifugation, 50 μl of the supernatant was retained as input, and the remaining part was incubated with Dynabeads M-280 Streptavidin (11205D, Thermo Fisher Scientific) mixture overnight at 4°C. .. The next day, a beads-probes-RNAs mixture was washed and incubated with 200 μl lysis buffer and proteinase K to reverse the formaldehyde crosslinking.

    Recombinant:

    Article Title: A polyomavirus peptide binds to the capsid VP1 pore and has potent antiviral activity against BK and JC polyomaviruses
    Article Snippet: Peptide-virus co-purification assay 54 nM peptide was incubated with 75 µg/mL (370 nM) recombinant full-length VP1 pentamers, or molar equivalent of VP1 pentamers of BK VLP or purified infectious BKV (as assessed by SDS-PAGE and comparing VP1 abundance by Coomassie stain) in 336 μl PBS buffer containing 1% DMSO and 0.01% Tween-20 for 1 hr at room temperature. .. 300 μl of mix was added to a fresh tube, and 20 μl of Dynabeads M-280 Streptavidin (Invitrogen) were added.

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  • 99
    Thermo Fisher streptavidin conjugated dynabeads
    Proteomic screen identifies ESCRT components as EphB2 interactors. (A) Strategy of purification and identification of the interactome of biotinylated EphB2 by mass spectrometry. (B) Representative images showing clustering of biotinylated FLAG-Avi-EphB2-YFP fusion protein around <t>streptavidin-conjugated</t> <t>Dynabeads</t> (within 5 min, right, stippled line). This effect required EphB2 biotinylation (left). (C) Western blot analysis showing tyrosine autophosphorylation (detected by anti-phospho EphB2 [Y594] or 4G10 antibodies) of biotinylated (BirA + ) but not unbiotinylated (BirA – ) FLAG-Avi-EphB2 in response to incubation with streptavidin-conjugated Dynabeads. Unfused Fc protein was used as negative control. Similar results were observed in three independent replicates. (D) Bar graph showing the SILAC ratios of representative members of different groups of the top 30 enriched full-length EphB2 interactors ( n = 3, mean ± SEM). Yellow, biotinylated versus unbiotinylated full-length EphB2 (FL EphB2); blue, biotinylated EphB2-ΔC versus unbiotinylated full-length EphB2 (EphB2-ΔC); gray, biotinylated full-length EphB2 versus biotinylated EphB2-ΔC (EphB2-cyto). (E) Full list of ESCRT complex components identified in the proteomic screen as interactors of full-length EphB2 in at least one of the three replicates. (F) Validation of the interaction between EphB2 and endogenous STAM1 or VPS4A by co-IP/Western blot (WB) analysis in HEK293 cells stably expressing EphB2. (G) Validation of the interaction between overexpressed EphB2 and STAM1 in HeLa cells. (H) Representative images showing that endogenous STAM and CHMP4B colocalize with surface EphB2 in HeLa cells. STAM and CHMP4B levels at the plasma membrane were increased 3.5- and 2-fold, respectively (highlighted by red triangles), in EphB2 + cells (indicated by yellow stippled line in the merge) compared with untransfected cells (white stippled line). Bars, 10 µm.
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    Proteomic screen identifies ESCRT components as EphB2 interactors. (A) Strategy of purification and identification of the interactome of biotinylated EphB2 by mass spectrometry. (B) Representative images showing clustering of biotinylated FLAG-Avi-EphB2-YFP fusion protein around streptavidin-conjugated Dynabeads (within 5 min, right, stippled line). This effect required EphB2 biotinylation (left). (C) Western blot analysis showing tyrosine autophosphorylation (detected by anti-phospho EphB2 [Y594] or 4G10 antibodies) of biotinylated (BirA + ) but not unbiotinylated (BirA – ) FLAG-Avi-EphB2 in response to incubation with streptavidin-conjugated Dynabeads. Unfused Fc protein was used as negative control. Similar results were observed in three independent replicates. (D) Bar graph showing the SILAC ratios of representative members of different groups of the top 30 enriched full-length EphB2 interactors ( n = 3, mean ± SEM). Yellow, biotinylated versus unbiotinylated full-length EphB2 (FL EphB2); blue, biotinylated EphB2-ΔC versus unbiotinylated full-length EphB2 (EphB2-ΔC); gray, biotinylated full-length EphB2 versus biotinylated EphB2-ΔC (EphB2-cyto). (E) Full list of ESCRT complex components identified in the proteomic screen as interactors of full-length EphB2 in at least one of the three replicates. (F) Validation of the interaction between EphB2 and endogenous STAM1 or VPS4A by co-IP/Western blot (WB) analysis in HEK293 cells stably expressing EphB2. (G) Validation of the interaction between overexpressed EphB2 and STAM1 in HeLa cells. (H) Representative images showing that endogenous STAM and CHMP4B colocalize with surface EphB2 in HeLa cells. STAM and CHMP4B levels at the plasma membrane were increased 3.5- and 2-fold, respectively (highlighted by red triangles), in EphB2 + cells (indicated by yellow stippled line in the merge) compared with untransfected cells (white stippled line). Bars, 10 µm.

    Journal: The Journal of Cell Biology

    Article Title: Exosomes mediate cell contact–independent ephrin-Eph signaling during axon guidance

    doi: 10.1083/jcb.201601085

    Figure Lengend Snippet: Proteomic screen identifies ESCRT components as EphB2 interactors. (A) Strategy of purification and identification of the interactome of biotinylated EphB2 by mass spectrometry. (B) Representative images showing clustering of biotinylated FLAG-Avi-EphB2-YFP fusion protein around streptavidin-conjugated Dynabeads (within 5 min, right, stippled line). This effect required EphB2 biotinylation (left). (C) Western blot analysis showing tyrosine autophosphorylation (detected by anti-phospho EphB2 [Y594] or 4G10 antibodies) of biotinylated (BirA + ) but not unbiotinylated (BirA – ) FLAG-Avi-EphB2 in response to incubation with streptavidin-conjugated Dynabeads. Unfused Fc protein was used as negative control. Similar results were observed in three independent replicates. (D) Bar graph showing the SILAC ratios of representative members of different groups of the top 30 enriched full-length EphB2 interactors ( n = 3, mean ± SEM). Yellow, biotinylated versus unbiotinylated full-length EphB2 (FL EphB2); blue, biotinylated EphB2-ΔC versus unbiotinylated full-length EphB2 (EphB2-ΔC); gray, biotinylated full-length EphB2 versus biotinylated EphB2-ΔC (EphB2-cyto). (E) Full list of ESCRT complex components identified in the proteomic screen as interactors of full-length EphB2 in at least one of the three replicates. (F) Validation of the interaction between EphB2 and endogenous STAM1 or VPS4A by co-IP/Western blot (WB) analysis in HEK293 cells stably expressing EphB2. (G) Validation of the interaction between overexpressed EphB2 and STAM1 in HeLa cells. (H) Representative images showing that endogenous STAM and CHMP4B colocalize with surface EphB2 in HeLa cells. STAM and CHMP4B levels at the plasma membrane were increased 3.5- and 2-fold, respectively (highlighted by red triangles), in EphB2 + cells (indicated by yellow stippled line in the merge) compared with untransfected cells (white stippled line). Bars, 10 µm.

    Article Snippet: 30 µl of streptavidin-conjugated Dynabeads (Dynabeads M-280 Streptavidin; Thermo Fisher Scientific) were added per dish to induce EphB2 clustering.

    Techniques: Purification, Mass Spectrometry, Western Blot, Incubation, Negative Control, Co-Immunoprecipitation Assay, Stable Transfection, Expressing

    Aβ22-41 binds to fibrinogen and fragment D. (A-B) Biotin-labeled Aβ42, Aβ1-16, Aβ15-25, and Aβ22-41 were incubated with fibrinogen (FBG) or fragment D (FD), and pulldown assays were carried out using streptavidin-coated

    Journal: Blood

    Article Title: Biochemical and structural analysis of the interaction between β-amyloid and fibrinogen

    doi: 10.1182/blood-2016-03-705228

    Figure Lengend Snippet: Aβ22-41 binds to fibrinogen and fragment D. (A-B) Biotin-labeled Aβ42, Aβ1-16, Aβ15-25, and Aβ22-41 were incubated with fibrinogen (FBG) or fragment D (FD), and pulldown assays were carried out using streptavidin-coated

    Article Snippet: Streptavidin-coated magnetic beads (Dynabeads M-280; Thermo-Fisher) were added for 30 minutes, washed, and eluted with nonreducing 1× lithium dodecyl sulfate sample buffer (Thermo Fisher Scientific).

    Techniques: Labeling, Incubation

    (a) Western blot analysis of CTB-, AV- and ST-bound MSC EVs. MSC CM was incubated with CTB, AV or ST followed by incubation with Dynabeads conjugated with Streptavidin. The beads were immobilised with a magnet, washed, denatured and resolved onto polyacrylamide gels before electroblotting onto a nitrocellulose membrane. The membrane was probed with a primary antibody followed by horseradish peroxidase-coupled secondary antibodies against the primary antibody. The blot was then incubated with a chemiluminescent HRP substrate to detect bound primary antibody. (b) 10 µg MSC EV was extracted sequentially with biotinylated CTB and then biotinylated AV or vice versa. After each extraction, the ligand-bound vesicles were removed with Dynabeads ® MyOne Streptavidin T1 and assayed for CD81 by ELISA. The relative level of CD81 in CTB-vesicles before and after extraction with AV, and that in AV-vesicles before and after extraction with CTB were normalized to that in AV-vesicles before CTB extraction. (c) RNA analysis of CTB-, AV- and ST-EVs. CTB-, AV- or ST-binding EVs were isolated as described above and extracted for RNA using Trizol. The pellet in each of extracts was re-suspended in 50 µL of RNase-free water. 10 µL of each RNA solution was resolved on a 15% Novex Tris-borate-EDTA(TBE)-urea gel before staining with ethidium bromide.

    Journal: Journal of Extracellular Vesicles

    Article Title: MSC secretes at least 3 EV types each with a unique permutation of membrane lipid, protein and RNA

    doi: 10.3402/jev.v5.29828

    Figure Lengend Snippet: (a) Western blot analysis of CTB-, AV- and ST-bound MSC EVs. MSC CM was incubated with CTB, AV or ST followed by incubation with Dynabeads conjugated with Streptavidin. The beads were immobilised with a magnet, washed, denatured and resolved onto polyacrylamide gels before electroblotting onto a nitrocellulose membrane. The membrane was probed with a primary antibody followed by horseradish peroxidase-coupled secondary antibodies against the primary antibody. The blot was then incubated with a chemiluminescent HRP substrate to detect bound primary antibody. (b) 10 µg MSC EV was extracted sequentially with biotinylated CTB and then biotinylated AV or vice versa. After each extraction, the ligand-bound vesicles were removed with Dynabeads ® MyOne Streptavidin T1 and assayed for CD81 by ELISA. The relative level of CD81 in CTB-vesicles before and after extraction with AV, and that in AV-vesicles before and after extraction with CTB were normalized to that in AV-vesicles before CTB extraction. (c) RNA analysis of CTB-, AV- and ST-EVs. CTB-, AV- or ST-binding EVs were isolated as described above and extracted for RNA using Trizol. The pellet in each of extracts was re-suspended in 50 µL of RNase-free water. 10 µL of each RNA solution was resolved on a 15% Novex Tris-borate-EDTA(TBE)-urea gel before staining with ethidium bromide.

    Article Snippet: The CTB, AV or ST reaction mix was added to 30 µL equivalent of Dynabeads M280 Streptavidin (Thermo Fisher Scientific, Waltham, MA) that were pre-washed as per manufacturer's instruction.

    Techniques: Western Blot, CtB Assay, Incubation, Enzyme-linked Immunosorbent Assay, Binding Assay, Isolation, Staining

    (a) SDS-PAGE analysis of MSC EVs extracted with membrane lipid-binding ligands, CTB, AV and ST, respectively. MSC CM was incubated with CTB, AV or ST followed by incubation with Dynabeads conjugated with Streptavidin. The beads were immobilized with a magnet and the supernatant was collected as the “unbound” fraction. The beads were then washed twice and the wash solutions were collected as “wash 1” and “wash 2,” respectively. The beads were re-suspended in PBS as the “bound” fraction. The equivalent of 20% of the starting samples (input) and each of their respective “unbound,” “wash 1,” “wash 2” and “bound” fractions were resolved onto polyacrylamide gels and the gels were stained with silver. (b) Size distribution of MSC EVs by NanoSight. MSC EVs were diluted 1,000× with 0.22 µm filtered PBS. The size distribution of exosome was then measured using NanoSight LM10 and analysed by Nanoparticles Tracking Analysis software according to the manufacturer's protocol. (c) SEM analysis of MSC EVs that were extracted with CTB, AV and ST. MSC EV preparation was incubated with biotinylated CTB, AV, ST or without ligand and then streptavidin-coated polystyrene particles. The beads were then washed twice with PBS and resuspended in PBS before being spotted and left to dry onto carbon tape on aluminium stubs at 40°C. The stubs were sputter coated with 2 ηm of gold coating (Leica Biosystems) and imaged in a Jeol 6701FESEM. Scale bar=100 ηm.

    Journal: Journal of Extracellular Vesicles

    Article Title: MSC secretes at least 3 EV types each with a unique permutation of membrane lipid, protein and RNA

    doi: 10.3402/jev.v5.29828

    Figure Lengend Snippet: (a) SDS-PAGE analysis of MSC EVs extracted with membrane lipid-binding ligands, CTB, AV and ST, respectively. MSC CM was incubated with CTB, AV or ST followed by incubation with Dynabeads conjugated with Streptavidin. The beads were immobilized with a magnet and the supernatant was collected as the “unbound” fraction. The beads were then washed twice and the wash solutions were collected as “wash 1” and “wash 2,” respectively. The beads were re-suspended in PBS as the “bound” fraction. The equivalent of 20% of the starting samples (input) and each of their respective “unbound,” “wash 1,” “wash 2” and “bound” fractions were resolved onto polyacrylamide gels and the gels were stained with silver. (b) Size distribution of MSC EVs by NanoSight. MSC EVs were diluted 1,000× with 0.22 µm filtered PBS. The size distribution of exosome was then measured using NanoSight LM10 and analysed by Nanoparticles Tracking Analysis software according to the manufacturer's protocol. (c) SEM analysis of MSC EVs that were extracted with CTB, AV and ST. MSC EV preparation was incubated with biotinylated CTB, AV, ST or without ligand and then streptavidin-coated polystyrene particles. The beads were then washed twice with PBS and resuspended in PBS before being spotted and left to dry onto carbon tape on aluminium stubs at 40°C. The stubs were sputter coated with 2 ηm of gold coating (Leica Biosystems) and imaged in a Jeol 6701FESEM. Scale bar=100 ηm.

    Article Snippet: The CTB, AV or ST reaction mix was added to 30 µL equivalent of Dynabeads M280 Streptavidin (Thermo Fisher Scientific, Waltham, MA) that were pre-washed as per manufacturer's instruction.

    Techniques: SDS Page, Binding Assay, CtB Assay, Incubation, Staining, Software