streptavidin agarose beads  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier
Bioz Manufacturer Symbol Thermo Fisher manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Name:
    Agarose
    Description:

    Catalog Number:
    75510019
    Price:
    None
    Category:
    Lab Reagents and Chemicals
    Buy from Supplier


    Structured Review

    Thermo Fisher streptavidin agarose beads
    Influence of phosphorylation on internalization of cell surface GFP-oatp1a1 in stably transfected HuH7 cells. HuH7-derived cell lines stably expressing GFP-oatp1a1 AA or GFP-oatp1a1 EE were prepared. These cells constitutively express PDZK1. Cells were surface biotinylated with membrane-impermeant sulfo-NHS-SS-biotin for 30 min at 4°C and then incubated at 37°C for up to 120 min to allow internalization. After removal of residual biotin from the cell surface by reduction, internalized biotinylated GFP-oatp1a1 was collected on <t>streptavidin-agarose</t> beads and subjected to immunoblot for oatp1a1. A : representative experiment. B : results of densitometric quantitation of 4 individual experiments. Data were normalized to total starting cell surface biotinylated oatp1a1. Lines are drawn through means at each time. Open symbols, oatp1a1 AA ; filled symbols, oatp1a1 EE .

    https://www.bioz.com/result/streptavidin agarose beads/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin agarose beads - by Bioz Stars, 2021-05
    97/100 stars

    Images

    1) Product Images from "PDZK1 binding and serine phosphorylation regulate subcellular trafficking of organic anion transport protein 1a1"

    Article Title: PDZK1 binding and serine phosphorylation regulate subcellular trafficking of organic anion transport protein 1a1

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00500.2010

    Influence of phosphorylation on internalization of cell surface GFP-oatp1a1 in stably transfected HuH7 cells. HuH7-derived cell lines stably expressing GFP-oatp1a1 AA or GFP-oatp1a1 EE were prepared. These cells constitutively express PDZK1. Cells were surface biotinylated with membrane-impermeant sulfo-NHS-SS-biotin for 30 min at 4°C and then incubated at 37°C for up to 120 min to allow internalization. After removal of residual biotin from the cell surface by reduction, internalized biotinylated GFP-oatp1a1 was collected on streptavidin-agarose beads and subjected to immunoblot for oatp1a1. A : representative experiment. B : results of densitometric quantitation of 4 individual experiments. Data were normalized to total starting cell surface biotinylated oatp1a1. Lines are drawn through means at each time. Open symbols, oatp1a1 AA ; filled symbols, oatp1a1 EE .
    Figure Legend Snippet: Influence of phosphorylation on internalization of cell surface GFP-oatp1a1 in stably transfected HuH7 cells. HuH7-derived cell lines stably expressing GFP-oatp1a1 AA or GFP-oatp1a1 EE were prepared. These cells constitutively express PDZK1. Cells were surface biotinylated with membrane-impermeant sulfo-NHS-SS-biotin for 30 min at 4°C and then incubated at 37°C for up to 120 min to allow internalization. After removal of residual biotin from the cell surface by reduction, internalized biotinylated GFP-oatp1a1 was collected on streptavidin-agarose beads and subjected to immunoblot for oatp1a1. A : representative experiment. B : results of densitometric quantitation of 4 individual experiments. Data were normalized to total starting cell surface biotinylated oatp1a1. Lines are drawn through means at each time. Open symbols, oatp1a1 AA ; filled symbols, oatp1a1 EE .

    Techniques Used: Stable Transfection, Transfection, Derivative Assay, Expressing, Incubation, Quantitation Assay

    Influence of phosphorylation on internalization of cell surface oatp1a1 in overnight-cultured rat hepatocytes. Hepatocytes were isolated from rat liver and cultured overnight, surface biotinylated with membrane-impermeant sulfo-NHS-SS-biotin for 30 min at 4°C, and then incubated at 37°C for 10 or 30 min in the absence (−) or presence (+) of 1 mM ATP. Previous studies showed that this short incubation of rat hepatocytes in extracellular ATP stimulates serine phosphorylation of oatp1a1 via activity of a purinergic receptor. After removal of residual biotin from the cell surface by reduction, internalized biotinylated oatp1a1 was collected on streptavidin-agarose beads and subjected to immunoblot for oatp1a1. A : representative study. B : densitometric quantitation of 3 experiments. Data were normalized to total starting cell surface biotinylated oatp1a1. Values are means ± SE.
    Figure Legend Snippet: Influence of phosphorylation on internalization of cell surface oatp1a1 in overnight-cultured rat hepatocytes. Hepatocytes were isolated from rat liver and cultured overnight, surface biotinylated with membrane-impermeant sulfo-NHS-SS-biotin for 30 min at 4°C, and then incubated at 37°C for 10 or 30 min in the absence (−) or presence (+) of 1 mM ATP. Previous studies showed that this short incubation of rat hepatocytes in extracellular ATP stimulates serine phosphorylation of oatp1a1 via activity of a purinergic receptor. After removal of residual biotin from the cell surface by reduction, internalized biotinylated oatp1a1 was collected on streptavidin-agarose beads and subjected to immunoblot for oatp1a1. A : representative study. B : densitometric quantitation of 3 experiments. Data were normalized to total starting cell surface biotinylated oatp1a1. Values are means ± SE.

    Techniques Used: Cell Culture, Isolation, Incubation, Activity Assay, Quantitation Assay

    2) Product Images from "Nitrosothiol-Trapping-Based Proteomic Analysis of S-Nitrosylation in Human Lung Carcinoma Cells"

    Article Title: Nitrosothiol-Trapping-Based Proteomic Analysis of S-Nitrosylation in Human Lung Carcinoma Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0169862

    SNO trapping-based analysis of S-nitrosylation in A549 cells. ( A) Schematic of the proteomic approach. Digitonin cell lysates, obtained from A549 treated with NO donor or with cytokines are incubated with a thioredoxin (Trx) trap mutant, Trx(C35S). In the trap mutant the resolving cysteine is replaced by serine (-OH). The protein also contains a streptavidin binding peptide. Trx(C35S) forms mixed disulfide bonds with nitrosylated substrates and the resulting complexes are pulled-down using avidin agarose. Identification of nitrosylation sites is assisted by differential thiol labeling, involving the sequential application of N-ethylmaleimide (NEM) and iodoacetamide (IAM). Proteins captured in the Trx pull-down are analyzed by SDS-PAGE or liquid chromatography-tandem mass spectrometry (LC-MS/MS). (B) A549 cells were treated with or without 500 μM S-nitrosocysteine (CysNO) for 10 min and thereafter digitonin lysates were incubated with Trx(C35S). Proteins captured by Trx were released by DTT and then analyzed by SDS-PAGE. Gels were stained with Krypton fluorescent protein stain and visualized using the Odyssey infrared imaging system. (C) A549 cells were treated for 72 h with LPS (0.5 mg/ml) and a cytokine mixture that included TNFα (20 ng/ml), IFN-γ (10 ng/ml) and IL-1β (10 ng/ml). Trx-based trapping of nitrosylated proteins was performed as in B.
    Figure Legend Snippet: SNO trapping-based analysis of S-nitrosylation in A549 cells. ( A) Schematic of the proteomic approach. Digitonin cell lysates, obtained from A549 treated with NO donor or with cytokines are incubated with a thioredoxin (Trx) trap mutant, Trx(C35S). In the trap mutant the resolving cysteine is replaced by serine (-OH). The protein also contains a streptavidin binding peptide. Trx(C35S) forms mixed disulfide bonds with nitrosylated substrates and the resulting complexes are pulled-down using avidin agarose. Identification of nitrosylation sites is assisted by differential thiol labeling, involving the sequential application of N-ethylmaleimide (NEM) and iodoacetamide (IAM). Proteins captured in the Trx pull-down are analyzed by SDS-PAGE or liquid chromatography-tandem mass spectrometry (LC-MS/MS). (B) A549 cells were treated with or without 500 μM S-nitrosocysteine (CysNO) for 10 min and thereafter digitonin lysates were incubated with Trx(C35S). Proteins captured by Trx were released by DTT and then analyzed by SDS-PAGE. Gels were stained with Krypton fluorescent protein stain and visualized using the Odyssey infrared imaging system. (C) A549 cells were treated for 72 h with LPS (0.5 mg/ml) and a cytokine mixture that included TNFα (20 ng/ml), IFN-γ (10 ng/ml) and IL-1β (10 ng/ml). Trx-based trapping of nitrosylated proteins was performed as in B.

    Techniques Used: Incubation, Mutagenesis, Binding Assay, Avidin-Biotin Assay, Labeling, SDS Page, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Staining, Imaging

    3) Product Images from "A Tyrosine Motif in the Cytoplasmic Domain of Mason-Pfizer Monkey Virus Is Essential for the Incorporation of Glycoprotein into Virions"

    Article Title: A Tyrosine Motif in the Cytoplasmic Domain of Mason-Pfizer Monkey Virus Is Essential for the Incorporation of Glycoprotein into Virions

    Journal: Journal of Virology

    doi: 10.1128/JVI.77.9.5192-5200.2003

    Biotinylation of envelope glycoproteins expressed on the cell surface. Two sets of cells expressing radiolabeled glycoprotein were biotinylated and immunoprecipitated with goat anti-M-PMV serum. (A) One set of immunoprecipitated samples was analyzed by SDS-PAGE. The mutant designation is shown above each lane, and the positions of the viral bands are indicated to the left. (B) The other set of immunoprecipitated samples was boiled in SDS, and streptavidin-agarose beads were added to isolated biotinylated Env proteins. The biotinylated Env proteins were then analyzed by SDS-PAGE. The positions of the viral bands are indicated to the left.
    Figure Legend Snippet: Biotinylation of envelope glycoproteins expressed on the cell surface. Two sets of cells expressing radiolabeled glycoprotein were biotinylated and immunoprecipitated with goat anti-M-PMV serum. (A) One set of immunoprecipitated samples was analyzed by SDS-PAGE. The mutant designation is shown above each lane, and the positions of the viral bands are indicated to the left. (B) The other set of immunoprecipitated samples was boiled in SDS, and streptavidin-agarose beads were added to isolated biotinylated Env proteins. The biotinylated Env proteins were then analyzed by SDS-PAGE. The positions of the viral bands are indicated to the left.

    Techniques Used: Expressing, Immunoprecipitation, SDS Page, Mutagenesis, Isolation

    4) Product Images from "Plasmodium falciparum Heterochromatin Protein 1 Marks Genomic Loci Linked to Phenotypic Variation of Exported Virulence Factors"

    Article Title: Plasmodium falciparum Heterochromatin Protein 1 Marks Genomic Loci Linked to Phenotypic Variation of Exported Virulence Factors

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1000569

    PfHP1 binds specifically to H3K9me3 and is associated with parasite heterochromatin. (A) Peptide competition demonstrates the specific binding of PfHP1 to H3K9me3. Recombinant PfHP1-HIS bound to a biotinylated H3K9me3 peptide was immobilized on streptavidin agarose beads. PfHP1 was only eluted by competition with the H3K9me3 peptide (lane 3), whereas peptides H3K9me3S10p (lane 5), H3K9ac (lane 7) and H4K20me3 (lane 11) were unable to compete. PfHP1 had weak affinity for H3K27me3 (lane 9). After elution with peptides, remaining PfHP1-HIS was eluted with high salt (lanes 4, 6, 8, 10, 12). P, peptide elution; S, high salt elution; FT, Flow-through after coupling of PfHP1-HIS; wash, last wash prior to peptide elution. The Western blot was probed with anti-6×HIS antibodies. (B) Solubility of PfHP1, H3, H4, gDNA and RNA after treatment of parasite nulcei with nucleases. Isolated nuclei were extracted either with low-salt, MNAse, DNAse or RNAse, followed by serial treatment of the insoluble pellets with high-salt and SDS. Lanes 1–4: PfHP1 and H3/H4 were tightly associated with the salt-insoluble fraction. Partially degraded RNA and undigested gDNA were apparent in the high-salt and SDS-soluble fractions. Lanes 5–8: Digestion with MNAse solubilizes approx. 50% of PfHP1 and the entire pool of H3/H4. RNA was degraded by the single-stranded nuclease activity of MNAse. Mononucleosomes (MN) were completely extracted with 1 M KCl. Lanes 9–12: After digestion of nuclei with DNAseI, PfHP1 and H3/H4 extracted equally with high-salt and SDS. Genomic DNA was completely digested and partially degraded RNA extracted with high-salt and SDS. Lanes 13–16: Treatment with RNAse A did not affect the extractablity of PfHP1 and H3/H4. RNA was completely digested by RNAse A whereas intact gDNA remained associated with the insoluble fraction. Equal amounts were analysed for each protein and nucleic acid sample. RNA in ethidium bromide stained gels was identified by re-examination of the gel after incubation in RNAse A-containing buffer for 1 hr at RT. W2, second wash after extraction with 1 M KCl.
    Figure Legend Snippet: PfHP1 binds specifically to H3K9me3 and is associated with parasite heterochromatin. (A) Peptide competition demonstrates the specific binding of PfHP1 to H3K9me3. Recombinant PfHP1-HIS bound to a biotinylated H3K9me3 peptide was immobilized on streptavidin agarose beads. PfHP1 was only eluted by competition with the H3K9me3 peptide (lane 3), whereas peptides H3K9me3S10p (lane 5), H3K9ac (lane 7) and H4K20me3 (lane 11) were unable to compete. PfHP1 had weak affinity for H3K27me3 (lane 9). After elution with peptides, remaining PfHP1-HIS was eluted with high salt (lanes 4, 6, 8, 10, 12). P, peptide elution; S, high salt elution; FT, Flow-through after coupling of PfHP1-HIS; wash, last wash prior to peptide elution. The Western blot was probed with anti-6×HIS antibodies. (B) Solubility of PfHP1, H3, H4, gDNA and RNA after treatment of parasite nulcei with nucleases. Isolated nuclei were extracted either with low-salt, MNAse, DNAse or RNAse, followed by serial treatment of the insoluble pellets with high-salt and SDS. Lanes 1–4: PfHP1 and H3/H4 were tightly associated with the salt-insoluble fraction. Partially degraded RNA and undigested gDNA were apparent in the high-salt and SDS-soluble fractions. Lanes 5–8: Digestion with MNAse solubilizes approx. 50% of PfHP1 and the entire pool of H3/H4. RNA was degraded by the single-stranded nuclease activity of MNAse. Mononucleosomes (MN) were completely extracted with 1 M KCl. Lanes 9–12: After digestion of nuclei with DNAseI, PfHP1 and H3/H4 extracted equally with high-salt and SDS. Genomic DNA was completely digested and partially degraded RNA extracted with high-salt and SDS. Lanes 13–16: Treatment with RNAse A did not affect the extractablity of PfHP1 and H3/H4. RNA was completely digested by RNAse A whereas intact gDNA remained associated with the insoluble fraction. Equal amounts were analysed for each protein and nucleic acid sample. RNA in ethidium bromide stained gels was identified by re-examination of the gel after incubation in RNAse A-containing buffer for 1 hr at RT. W2, second wash after extraction with 1 M KCl.

    Techniques Used: Binding Assay, Recombinant, Flow Cytometry, Western Blot, Solubility, Isolation, Activity Assay, Staining, Incubation

    5) Product Images from "Expanding the Functional Repertoire of CTD Kinase I and RNA Polymerase II: Novel PhosphoCTD-Associating Proteins in the Yeast Proteome †"

    Article Title: Expanding the Functional Repertoire of CTD Kinase I and RNA Polymerase II: Novel PhosphoCTD-Associating Proteins in the Yeast Proteome †

    Journal: Biochemistry

    doi: 10.1021/bi048364h

    Specificity of binding of various PCAPs to (phospho)CTD peptides using BIACORE (surface plasmon resonance) analysis. (A) Sequences of biotinylated peptides that were immobilized on streptavidin sensor chips with symbols used in graphs B–J. (B) Sensorgrams of binding of MBP–Set2(425–551) (2 µ M) to the 2,5P, 5P, and NP CTD peptides, after subtracting signal from an empty flow cell (nonspecific binding). (C) Sensorgrams of binding of MBP–Ssd1(1–160) (2 µ M) to CTD peptides as in (B). (D) Sensorgrams of binding of Prp40 (cFF domains) [amino acids 267–583 ( 34 )] (2 µ M) to CTD peptides as in (B). (E) Sensorgrams of binding of MBP–Hrr25(1– 494) (2 µ M) to CTD peptides as in (B). (F) Sensorgrams of binding Ess1 (WW domain) (2 µ M) to CTD peptides as in (B). (G) Sensorgrams of binding of MBP–Ssd1(1–160) (2 µ M) to the 2,5P, 5P, and 2P CTD peptides, after subtracting signal from the scrambled 6PC peptide channel [sequences of peptides shown in (A)]. (H) Sensorgrams of binding of Prp40 (cFF domains) [amino acids 267–583 ( 34 )] (2 µ M) to CTD peptides as in (G). (I) Sensorgrams of binding of MBP–Hrr25(1–494) (2 µ M) to CTD peptides as in (G). (J) Sensorgrams of binding of Ess1 (WW domain) (2 µ M) to CTD peptides as in (G).
    Figure Legend Snippet: Specificity of binding of various PCAPs to (phospho)CTD peptides using BIACORE (surface plasmon resonance) analysis. (A) Sequences of biotinylated peptides that were immobilized on streptavidin sensor chips with symbols used in graphs B–J. (B) Sensorgrams of binding of MBP–Set2(425–551) (2 µ M) to the 2,5P, 5P, and NP CTD peptides, after subtracting signal from an empty flow cell (nonspecific binding). (C) Sensorgrams of binding of MBP–Ssd1(1–160) (2 µ M) to CTD peptides as in (B). (D) Sensorgrams of binding of Prp40 (cFF domains) [amino acids 267–583 ( 34 )] (2 µ M) to CTD peptides as in (B). (E) Sensorgrams of binding of MBP–Hrr25(1– 494) (2 µ M) to CTD peptides as in (B). (F) Sensorgrams of binding Ess1 (WW domain) (2 µ M) to CTD peptides as in (B). (G) Sensorgrams of binding of MBP–Ssd1(1–160) (2 µ M) to the 2,5P, 5P, and 2P CTD peptides, after subtracting signal from the scrambled 6PC peptide channel [sequences of peptides shown in (A)]. (H) Sensorgrams of binding of Prp40 (cFF domains) [amino acids 267–583 ( 34 )] (2 µ M) to CTD peptides as in (G). (I) Sensorgrams of binding of MBP–Hrr25(1–494) (2 µ M) to CTD peptides as in (G). (J) Sensorgrams of binding of Ess1 (WW domain) (2 µ M) to CTD peptides as in (G).

    Techniques Used: Binding Assay, SPR Assay, Flow Cytometry

    6) Product Images from "Entamoeba histolytica Cell Surface Calreticulin Binds Human C1q and Functions in Amebic Phagocytosis of Host Cells"

    Article Title: Entamoeba histolytica Cell Surface Calreticulin Binds Human C1q and Functions in Amebic Phagocytosis of Host Cells

    Journal: Infection and Immunity

    doi: 10.1128/IAI.06287-11

    Calreticulin is also present on the surface of unstimulated trophozoites. Amebic trophozoites were incubated with or without healthy Jurkat lymphocytes (37°C for 5 min), and then surface proteins were specifically biotinylated with sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated using streptavidin-coated beads and analyzed by Western blotting using antibodies specific for the GalNAc lectin (positive control), the cytosolic protein ICP1 (negative control), and calreticulin. Nonbiotinylated whole E. histolytica lysates were run as a positive control for the Western blot.
    Figure Legend Snippet: Calreticulin is also present on the surface of unstimulated trophozoites. Amebic trophozoites were incubated with or without healthy Jurkat lymphocytes (37°C for 5 min), and then surface proteins were specifically biotinylated with sulfo-NHS-LC-biotin. Biotinylated proteins were precipitated using streptavidin-coated beads and analyzed by Western blotting using antibodies specific for the GalNAc lectin (positive control), the cytosolic protein ICP1 (negative control), and calreticulin. Nonbiotinylated whole E. histolytica lysates were run as a positive control for the Western blot.

    Techniques Used: Incubation, Western Blot, Positive Control, Negative Control

    E. histolytica calreticulin binds to apoptotic lymphocytes and to human C1q. (A) Calreticulin binds specifically to apoptotic lymphocytes, independent of C1q. Healthy and apoptotic Jurkat lymphocytes cultured in serum-free medium were stained for flow cytometry using biotin-calreticulin or biotin-BSA (negative control) and streptavidin-Alexa Fluor 488. Representative histograms of amebic fluorescence are shown. (B) Calreticulin binds directly to human C1q. Recombinant calreticulin (rCalreticulin) binding to immobilized C1q was measured by ELISA using biotinylated anti-calreticulin scFv and streptavidin-HRP. The graph shows a single-site specific binding curve for binding of the indicated concentration of calreticulin, generated by nonlinear regression (mean and SD; n = 3). (C) Specific inhibition of calreticulin-C1q binding by free C1q and collectins. Binding assays were performed in the presence of 3-fold molar excess BSA (negative control), C1q, MBL, and SP-A. Calreticulin binding was expressed as the percentage of specific binding measured in buffer alone (mean and SD; n = 3). *, P
    Figure Legend Snippet: E. histolytica calreticulin binds to apoptotic lymphocytes and to human C1q. (A) Calreticulin binds specifically to apoptotic lymphocytes, independent of C1q. Healthy and apoptotic Jurkat lymphocytes cultured in serum-free medium were stained for flow cytometry using biotin-calreticulin or biotin-BSA (negative control) and streptavidin-Alexa Fluor 488. Representative histograms of amebic fluorescence are shown. (B) Calreticulin binds directly to human C1q. Recombinant calreticulin (rCalreticulin) binding to immobilized C1q was measured by ELISA using biotinylated anti-calreticulin scFv and streptavidin-HRP. The graph shows a single-site specific binding curve for binding of the indicated concentration of calreticulin, generated by nonlinear regression (mean and SD; n = 3). (C) Specific inhibition of calreticulin-C1q binding by free C1q and collectins. Binding assays were performed in the presence of 3-fold molar excess BSA (negative control), C1q, MBL, and SP-A. Calreticulin binding was expressed as the percentage of specific binding measured in buffer alone (mean and SD; n = 3). *, P

    Techniques Used: Cell Culture, Staining, Flow Cytometry, Cytometry, Negative Control, Fluorescence, Recombinant, Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Generated, Inhibition

    7) Product Images from "Intracellular Na+ Controls Cell Surface Expression of Na,K-ATPase via a cAMP-independent PKA Pathway in Mammalian Kidney Collecting Duct Cells"

    Article Title: Intracellular Na+ Controls Cell Surface Expression of Na,K-ATPase via a cAMP-independent PKA Pathway in Mammalian Kidney Collecting Duct Cells

    Journal: Molecular Biology of the Cell

    doi: 10.1091/mbc.E02-11-0720

    Effect of high [Na + ]i on Na,K-ATPase cell surface and total expression in mpkCCD c14 cells. Confluent mpkCCDcl4 cells grown on polycarbonate filters were first preincubated in the presence of 15 or 135 mM Na + for 30 min at 37°C and then permeabilized by 1 μg/ml amphotericin B or 0.1 U/μl nystatin, or incubated in K + -free medium for 1 h at 37°C. The Na,K-ATPase α-subunit was detected by Western blotting performed after biotinylation and streptavidin precipitation of cell surface proteins (Cell surface Na,K-ATPase; A and B) or on cell lysates (Total Na,K-ATPase; C and D). (A and C) Representative immunoblots showing cell surface (A) and total (C) expression of Na,K-ATPase. (B and D) Bars represent densitometric values expressed as the percentage of the optical density value measured in the presence of 15 mM Na + . Results are means ± SE from five independent experiments. *p
    Figure Legend Snippet: Effect of high [Na + ]i on Na,K-ATPase cell surface and total expression in mpkCCD c14 cells. Confluent mpkCCDcl4 cells grown on polycarbonate filters were first preincubated in the presence of 15 or 135 mM Na + for 30 min at 37°C and then permeabilized by 1 μg/ml amphotericin B or 0.1 U/μl nystatin, or incubated in K + -free medium for 1 h at 37°C. The Na,K-ATPase α-subunit was detected by Western blotting performed after biotinylation and streptavidin precipitation of cell surface proteins (Cell surface Na,K-ATPase; A and B) or on cell lysates (Total Na,K-ATPase; C and D). (A and C) Representative immunoblots showing cell surface (A) and total (C) expression of Na,K-ATPase. (B and D) Bars represent densitometric values expressed as the percentage of the optical density value measured in the presence of 15 mM Na + . Results are means ± SE from five independent experiments. *p

    Techniques Used: Expressing, Incubation, Western Blot

    8) Product Images from "A vitronectin-derived peptide reverses ovariectomy-induced bone loss via regulation of osteoblast and osteoclast differentiation"

    Article Title: A vitronectin-derived peptide reverses ovariectomy-induced bone loss via regulation of osteoblast and osteoclast differentiation

    Journal: Cell Death and Differentiation

    doi: 10.1038/cdd.2017.153

    VnP-16 promotes osteogenic cell attachment through direct interaction with β 1 integrin. ( a ) Attachment of human osteogenic cells pretreated with EDTA (5 mM), MnCl 2 (500 μ M), or heparin (100 μ g/ml) to VnP-16. The cells were seeded onto plates that were precoated with VnP-16 (9.1 μ g/cm 2 ) for 1 h. ( b ) The effects of various integrin-blocking antibodies on cell attachment to VnP-16. ( c–e ) Immunoblotting ( c ) and densitometric analysis ( d ) of β 1 integrin, and cell attachment to VnP-16 ( e ) in osteogenic cells that were transfected with a control (Con) or β 1 integrin-specific siRNA (10 nM; β 1 integrin). ( f ) Streptavidin-bead pulldown assay with the biotinylated SP or biotinylated VnP-16 peptides from extracts of osteogenic cells that were cultured on biotinylated SP- or biotinylated VnP-16-coated dishes for 30 min. Data in ( a , b,e ) ( n =4), and ( d ) ( n =2) represent the mean±SD. ** P
    Figure Legend Snippet: VnP-16 promotes osteogenic cell attachment through direct interaction with β 1 integrin. ( a ) Attachment of human osteogenic cells pretreated with EDTA (5 mM), MnCl 2 (500 μ M), or heparin (100 μ g/ml) to VnP-16. The cells were seeded onto plates that were precoated with VnP-16 (9.1 μ g/cm 2 ) for 1 h. ( b ) The effects of various integrin-blocking antibodies on cell attachment to VnP-16. ( c–e ) Immunoblotting ( c ) and densitometric analysis ( d ) of β 1 integrin, and cell attachment to VnP-16 ( e ) in osteogenic cells that were transfected with a control (Con) or β 1 integrin-specific siRNA (10 nM; β 1 integrin). ( f ) Streptavidin-bead pulldown assay with the biotinylated SP or biotinylated VnP-16 peptides from extracts of osteogenic cells that were cultured on biotinylated SP- or biotinylated VnP-16-coated dishes for 30 min. Data in ( a , b,e ) ( n =4), and ( d ) ( n =2) represent the mean±SD. ** P

    Techniques Used: Cell Attachment Assay, Blocking Assay, Transfection, Cell Culture

    9) Product Images from "Morphine‐mediated release of miR‐138 in astrocyte‐derived extracellular vesicles promotes microglial activation, et al. Morphine‐mediated release of miR‐138 in astrocyte‐derived extracellular vesicles promotes microglial activation"

    Article Title: Morphine‐mediated release of miR‐138 in astrocyte‐derived extracellular vesicles promotes microglial activation, et al. Morphine‐mediated release of miR‐138 in astrocyte‐derived extracellular vesicles promotes microglial activation

    Journal: Journal of Extracellular Vesicles

    doi: 10.1002/jev2.12027

    ADEV miR‐138 interacts with murine TLR7 in the endosomes. (A) Representative fluorescence images of mouse primary microglial cells incubated with Exo‐Fect+ADEVs+Cy5‐miR138 or Exo‐Fect+Cy5‐miR138 (without ADEVs) or Exo‐Fect+ADEVs+miR138 (unstained) for 30 min followed by immunostaining of (A) an early endosome marker (EEA1, Green) and (B) TLR7 (Green). Exo‐fect+ADEVs+Cy5‐miR138, Cy5‐miR138 were loaded into ADEVs using by Exo‐Fect transfection kit; Exo‐fect+Cy5‐miR138, Cy5‐miR138 were loaded using by Exo‐Fect transfection kit (without ADEVs); Exo‐fect+ADEVs+miR138, unstained miR138 were loaded into ADEVs using Exo‐Fect transfection kit; Bars, 50 μm ( n = 3). (C) TLR7 was immunoprecipitated from BV2 cells by IgG / TLR7 / LAMP1 antibody, followed by assessment of miR138 / U6 expression by real‐time PCR. One‐way ANOVA followed by Bonferroni's post hoc test was used to determine the statistical significance among multiple groups ( n = 3). (D) TLR7 was immunoprecipitated from HEK‐Null / HEK‐TLR7 cells by IgG/TLR7 antibody, followed by assessment of miR138 expression by real‐time PCR. One‐way ANOVA followed by Bonferroni's post hoc test was used to determine the statistical significance among multiple groups ( n = 3). (E) The protein of TLR7 were pull down by miR‐138‐biotin / miR‐mut‐138‐biotin with Streptavidin agarose beads in BV2 cells. All data are presented as mean ± SD or SEM of three individual experiments. *, P
    Figure Legend Snippet: ADEV miR‐138 interacts with murine TLR7 in the endosomes. (A) Representative fluorescence images of mouse primary microglial cells incubated with Exo‐Fect+ADEVs+Cy5‐miR138 or Exo‐Fect+Cy5‐miR138 (without ADEVs) or Exo‐Fect+ADEVs+miR138 (unstained) for 30 min followed by immunostaining of (A) an early endosome marker (EEA1, Green) and (B) TLR7 (Green). Exo‐fect+ADEVs+Cy5‐miR138, Cy5‐miR138 were loaded into ADEVs using by Exo‐Fect transfection kit; Exo‐fect+Cy5‐miR138, Cy5‐miR138 were loaded using by Exo‐Fect transfection kit (without ADEVs); Exo‐fect+ADEVs+miR138, unstained miR138 were loaded into ADEVs using Exo‐Fect transfection kit; Bars, 50 μm ( n = 3). (C) TLR7 was immunoprecipitated from BV2 cells by IgG / TLR7 / LAMP1 antibody, followed by assessment of miR138 / U6 expression by real‐time PCR. One‐way ANOVA followed by Bonferroni's post hoc test was used to determine the statistical significance among multiple groups ( n = 3). (D) TLR7 was immunoprecipitated from HEK‐Null / HEK‐TLR7 cells by IgG/TLR7 antibody, followed by assessment of miR138 expression by real‐time PCR. One‐way ANOVA followed by Bonferroni's post hoc test was used to determine the statistical significance among multiple groups ( n = 3). (E) The protein of TLR7 were pull down by miR‐138‐biotin / miR‐mut‐138‐biotin with Streptavidin agarose beads in BV2 cells. All data are presented as mean ± SD or SEM of three individual experiments. *, P

    Techniques Used: Fluorescence, Incubation, Immunostaining, Marker, Transfection, Immunoprecipitation, Expressing, Real-time Polymerase Chain Reaction

    10) Product Images from "Novel chloroacetamido compound CWR-J02 is an anti-inflammatory glutaredoxin-1 inhibitor"

    Article Title: Novel chloroacetamido compound CWR-J02 is an anti-inflammatory glutaredoxin-1 inhibitor

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0187991

    J02 interactome for BV2 cells–proteins involved in inflammatory responses. BV2 cells were treated with 40 μM J02 or DMSO. Resulting cell pellets were lysed, linked to a biotin azide probe, and run over a streptavidin column. A , Pulled down proteins were identified using mass spectrometry (see SI Materials and Methods section for further details). A, Inflammatory proteins shown to be regulated via S-glutathionylation and identified in the mass spectrometry dataset (supplemental S1 Table ), including glutaredoxin-1. B , Grx1 and p65 are detected in J02-adducted samples. BV2 cells were treated with 40 μM J02 or equivalent volume DMSO for 30 min. Medium was changed, and cells were allowed to recover for 60 min. Resulting cell pellets were lysed, adducted with azide fluorescent fluorophore, and run over streptavidin beads to precipitate J02-adducted proteins. See Materials and Methods for further details. Eluted proteins were separated on SDS-PAGE gel, transferred to PVDF membrane, and probed with antibodies against Grx1 (left) and p65 (right).
    Figure Legend Snippet: J02 interactome for BV2 cells–proteins involved in inflammatory responses. BV2 cells were treated with 40 μM J02 or DMSO. Resulting cell pellets were lysed, linked to a biotin azide probe, and run over a streptavidin column. A , Pulled down proteins were identified using mass spectrometry (see SI Materials and Methods section for further details). A, Inflammatory proteins shown to be regulated via S-glutathionylation and identified in the mass spectrometry dataset (supplemental S1 Table ), including glutaredoxin-1. B , Grx1 and p65 are detected in J02-adducted samples. BV2 cells were treated with 40 μM J02 or equivalent volume DMSO for 30 min. Medium was changed, and cells were allowed to recover for 60 min. Resulting cell pellets were lysed, adducted with azide fluorescent fluorophore, and run over streptavidin beads to precipitate J02-adducted proteins. See Materials and Methods for further details. Eluted proteins were separated on SDS-PAGE gel, transferred to PVDF membrane, and probed with antibodies against Grx1 (left) and p65 (right).

    Techniques Used: Mass Spectrometry, SDS Page

    Related Articles

    Centrifugation:

    Article Title: The Purinergic Receptor P2X7 Triggers ?-Secretase-dependent Processing of the Amyloid Precursor Protein *
    Article Snippet: .. After centrifugation at 270,000 × g at 4 °C for 15 h, 2-ml aliquots of the gradients were collected from the top and submitted to precipitation with neutravidin-agarose beads (Pierce) at 4 °C overnight. .. Neutravidin-bound proteins from each sucrose gradient fraction were separated by SDS-PAGE and transferred to nitrocellulose membranes.

    Immunoprecipitation:

    Article Title: Vital function of PRELI and essential requirement of its LEA motif
    Article Snippet: For AIF/PRELI or PRELI/LEA− interactions, 1.0 mg V5-tagged PRELI or V5-tagged PRELI/LEA− protein lysates from transient HeLa and DOX-induced Blin-1 transfectants were immunoprecipitated with agarose-conjugated anti-AIF antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and blotted. .. For OPA1/PRELI or PRELI/LEA− interactions, 1.0 mg of the above cited protein lysates were immunoprecipitated with agarose-conjugated anti-V5 antibody (Invitrogen) and blotted. .. The blots were reacted with HRP-conjugated anti-V5 monoclonal antibody (Invitrogen) or anti-OPA1 monoclonal antibody followed by an HRP-conjugated anti-mouse IgG reaction.

    Article Title: Cystine 186-cystine 209 disulfide bond is not essential for the procoagulant activity of tissue factor or for its de-encryption
    Article Snippet: .. For immunoprecipitation, biotin-labeled cells (from 60-mm culture dish) were solubilized in buffer A containing 1% Triton X-100 (1.5 mL) supplemented with cocktail of protease inhibitors, and the cell lysates were incubated overnight at 4°C with high-capacity neutravidin-agarose resin (50 μL; Pierce Chemical). .. After the removal of the unbound material, the beads were washed 3 times with the washing buffer, and the bound material was eluted with 100 μL of 2× sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer.

    Purification:

    Article Title: De novo macrocyclic peptides for inhibiting, stabilising and probing the function of the Retromer endosomal trafficking complex
    Article Snippet: RT-L4 Binding Site Screening Assay Mapping the potential binding region of RT-L4 was carried out using purified Vps26A, Vps351-390 , Vps26-Vps351-390 subcomplex and the associated mutants. .. First, 10 μM of purified proteins were incubated with fresh streptavidin agarose containing either 100 μM of RT-L4 or equivalent percentage (v/v) of DMSO. .. The mixture was incubated in binding buffer containing 50 mM HEPES pH 7.5, 200 mM NaCl, 5% glycerol, 0.5 mM TCEP for 30 min at 4°C.

    Incubation:

    Article Title: De novo macrocyclic peptides for inhibiting, stabilising and probing the function of the Retromer endosomal trafficking complex
    Article Snippet: RT-L4 Binding Site Screening Assay Mapping the potential binding region of RT-L4 was carried out using purified Vps26A, Vps351-390 , Vps26-Vps351-390 subcomplex and the associated mutants. .. First, 10 μM of purified proteins were incubated with fresh streptavidin agarose containing either 100 μM of RT-L4 or equivalent percentage (v/v) of DMSO. .. The mixture was incubated in binding buffer containing 50 mM HEPES pH 7.5, 200 mM NaCl, 5% glycerol, 0.5 mM TCEP for 30 min at 4°C.

    Article Title: Metalloprotease ADAM10 Is Required for Notch1 Site 2 Cleavage *
    Article Snippet: .. After input, samples were taken, and streptavidin-coupled agarose beads (Pierce) were added to the supernatant and incubated for 90 min. .. Samples were washed three times in RIPA and three times in PBS, boiled in Laemmli buffer, and analyzed by PAGE.

    Article Title: Cystine 186-cystine 209 disulfide bond is not essential for the procoagulant activity of tissue factor or for its de-encryption
    Article Snippet: .. For immunoprecipitation, biotin-labeled cells (from 60-mm culture dish) were solubilized in buffer A containing 1% Triton X-100 (1.5 mL) supplemented with cocktail of protease inhibitors, and the cell lysates were incubated overnight at 4°C with high-capacity neutravidin-agarose resin (50 μL; Pierce Chemical). .. After the removal of the unbound material, the beads were washed 3 times with the washing buffer, and the bound material was eluted with 100 μL of 2× sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer.

    Molecular Weight:

    Article Title: Variant surface glycoproteins from Venezuelan trypanosome isolates are recognized by sera from animals infected with either Trypanosoma evansi or Trypanosoma vivax
    Article Snippet: In this study, we investigated the potential use of VSG variants as diagnostic reagents for the detection of trypanosomosis caused by T. vivax and T. evansi , and examined whether the soluble form of these VSG antigens contained common epitopes recognized by sera from animals infected with either of these species of trypanosomes. .. 2.1 Materials Reagents were purchased from the following sources: middle range molecular weight protein markers, anti-mouse IgG horseradish peroxidase conjugate, 5-bromo-4-chloro-3 indolyl phosphate (BCIP), nitro blue tetrazolium (NBT), Promega ; anti-rabbit IgG alkaline phosphatase conjugate, anti-bovine IgG horseradish peroxidase conjugate, anti-bovine IgG alkaline phosphatase conjugate, anti-equine IgG alkaline phosphatase conjugate, anti-mouse IgG alkaline phosphatase conjugate, diaminobenzidine (DAB), horseradish peroxidase type VI-A, fibrous DEAE-cellulose, benzamidine, iodoacetamide, phenyl methyl sulfonyl fluoride (PMSF), N-N′-1,2 phenylenedimaleimide (o-PDM), N-N′-1,4 phenyllenedimaleimide (p-PDM), gel filtration molecular weight protein marker kit, Staphylococcus aureus V8 protease, concanavalin A (Con A), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), methyl-α-d -mannopyranoside, methyl-α-d -glucopyranoside, Sigma ; Q-Sepharose, S-Sepharose, Sefacryl S-300, Pharmacia ; pre-stained high molecular weight protein markers, Gibco BRL ; sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC), nitrocellulose (0.45 μm pore size), Pierce; broad range isoelectric focusing calibration kit (3–10), Immobilin dry strips (pH 5–8), ampholites (pH 3–10), BioRad . ..

    Filtration:

    Article Title: Variant surface glycoproteins from Venezuelan trypanosome isolates are recognized by sera from animals infected with either Trypanosoma evansi or Trypanosoma vivax
    Article Snippet: In this study, we investigated the potential use of VSG variants as diagnostic reagents for the detection of trypanosomosis caused by T. vivax and T. evansi , and examined whether the soluble form of these VSG antigens contained common epitopes recognized by sera from animals infected with either of these species of trypanosomes. .. 2.1 Materials Reagents were purchased from the following sources: middle range molecular weight protein markers, anti-mouse IgG horseradish peroxidase conjugate, 5-bromo-4-chloro-3 indolyl phosphate (BCIP), nitro blue tetrazolium (NBT), Promega ; anti-rabbit IgG alkaline phosphatase conjugate, anti-bovine IgG horseradish peroxidase conjugate, anti-bovine IgG alkaline phosphatase conjugate, anti-equine IgG alkaline phosphatase conjugate, anti-mouse IgG alkaline phosphatase conjugate, diaminobenzidine (DAB), horseradish peroxidase type VI-A, fibrous DEAE-cellulose, benzamidine, iodoacetamide, phenyl methyl sulfonyl fluoride (PMSF), N-N′-1,2 phenylenedimaleimide (o-PDM), N-N′-1,4 phenyllenedimaleimide (p-PDM), gel filtration molecular weight protein marker kit, Staphylococcus aureus V8 protease, concanavalin A (Con A), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), methyl-α-d -mannopyranoside, methyl-α-d -glucopyranoside, Sigma ; Q-Sepharose, S-Sepharose, Sefacryl S-300, Pharmacia ; pre-stained high molecular weight protein markers, Gibco BRL ; sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC), nitrocellulose (0.45 μm pore size), Pierce; broad range isoelectric focusing calibration kit (3–10), Immobilin dry strips (pH 5–8), ampholites (pH 3–10), BioRad . ..

    Marker:

    Article Title: Variant surface glycoproteins from Venezuelan trypanosome isolates are recognized by sera from animals infected with either Trypanosoma evansi or Trypanosoma vivax
    Article Snippet: In this study, we investigated the potential use of VSG variants as diagnostic reagents for the detection of trypanosomosis caused by T. vivax and T. evansi , and examined whether the soluble form of these VSG antigens contained common epitopes recognized by sera from animals infected with either of these species of trypanosomes. .. 2.1 Materials Reagents were purchased from the following sources: middle range molecular weight protein markers, anti-mouse IgG horseradish peroxidase conjugate, 5-bromo-4-chloro-3 indolyl phosphate (BCIP), nitro blue tetrazolium (NBT), Promega ; anti-rabbit IgG alkaline phosphatase conjugate, anti-bovine IgG horseradish peroxidase conjugate, anti-bovine IgG alkaline phosphatase conjugate, anti-equine IgG alkaline phosphatase conjugate, anti-mouse IgG alkaline phosphatase conjugate, diaminobenzidine (DAB), horseradish peroxidase type VI-A, fibrous DEAE-cellulose, benzamidine, iodoacetamide, phenyl methyl sulfonyl fluoride (PMSF), N-N′-1,2 phenylenedimaleimide (o-PDM), N-N′-1,4 phenyllenedimaleimide (p-PDM), gel filtration molecular weight protein marker kit, Staphylococcus aureus V8 protease, concanavalin A (Con A), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), methyl-α-d -mannopyranoside, methyl-α-d -glucopyranoside, Sigma ; Q-Sepharose, S-Sepharose, Sefacryl S-300, Pharmacia ; pre-stained high molecular weight protein markers, Gibco BRL ; sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC), nitrocellulose (0.45 μm pore size), Pierce; broad range isoelectric focusing calibration kit (3–10), Immobilin dry strips (pH 5–8), ampholites (pH 3–10), BioRad . ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher streptavidin coated magnetic beads
    Dual-aptamer detection schematic Thrombin and PDGF-BB are represented by triangles and double-ellipses, respectively. The sensing aptamers (MB-T, MB-P) containing thrombin aptamer (Apt 1A) and PDGF-BB aptamer (Apt 2A) are each labeled with an MB tag (MB1 and MB2) and flanked by a pair of universal primers (5′p and 3′p). Capture aptamers (Bio-T and Bio-P) containing thrombin aptamer (Apt 1B) and PDGF-BB aptamer (Apt 2B) are biotinylated to be attached to <t>streptavidin-coated</t> magnetic beads. Oligo(dT)s are used as the spacer between the aptamer sequences and the beads. After separation, eluted sensing aptamers are amplified by multiplex PCR and quantified.
    Streptavidin Coated Magnetic Beads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin coated magnetic beads/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin coated magnetic beads - by Bioz Stars, 2021-05
    99/100 stars
      Buy from Supplier

    95
    Thermo Fisher streptavidin agarose beads
    Insulin-induced translocation of GLUT4 is disrupted in muscle fibers isolated from HTZ mice. ( a–c ) Freshly dissected FDB muscles from 2 month-old WT and HTZ mice were digested with collagenase. Isolated fibers were stimulated during 15 min with 0.1 µM insulin to induce GLUT4 translocation, fixed and immunolabeled with a polyclonal-GLUT4 antibody. Translocation of GLUT4 was estimated by measuring the total intensity fluorescence of GLUT4 in ROIs at the sarcolemma. ( a ) Examples of the ROIs used are drawn in white. GLUT4 was measured on both edges of the confocal image and then averaged. ( b ) Examples images of GLUT4 signal in WT and HTZ fibers at the resting (left panels) and insulin-stimulated condition (right panels). Scale bar = 20 µm. ( c ) The graph show the averaged GLUT4 signal in sarcolemma. Note that insulin-induced GLUT4 translocation is significantly reduced in HTZ myofibers compared to WT myofibers. Data are expressed as mean GLUT4 fluorescence signal ± SEM. Statistical comparisons were performed utilizing a two-tail t-test Welch corrected for parametric data. The symbols * and # denote significance with respect to WT-resting and WT-insulin-stimulated fibers, respectively. N is between 34 and 66 fibers from at least 10 different animals per genotype. ( d ) FDB muscles were dissected from WT and HTZ mice, stabilized in Tyrode solution, stimulated for 30 min with 0.1 µM insulin and then exposed to 1 mg/ml of biotin at 4 °C during 60 min. After quenching with 100 mM glycine, muscles were frozen and pulverized in liquid nitrogen, lysed and centrifuged at 14.000 g for 10 min. Supernatants were mixed with <t>streptavidin-agarose</t> beads overnight at 4 °C and then centrifuged at 14.000 g for 3 min. Biotinylated and non-biotinylated fractions were used to evaluate GLUT4 expression by western blot. GAPDH was used as a control that only surface proteins were labeled in biotinylated fractions. On the left are shown representative blots per each condition, on the right are plotted the percentages of GLUT4 in biotinylated fractions. Data are expressed as mean GLUT4% ± SEM. Statistical comparisons were performed utilizing a two-tail t-test Welch corrected for parametric data. The symbol * denote significance with respect to WT-muscles. N is five different animals per genotype.
    Streptavidin Agarose Beads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin agarose beads/product/Thermo Fisher
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin agarose beads - by Bioz Stars, 2021-05
    95/100 stars
      Buy from Supplier

    97
    Thermo Fisher streptavidin coupled agarose beads
    Notch1 S2 cleavage occurs at the cell surface. A, upper panel, Myc immunoblot of surface-biotinylated and <t>streptavidin-precipitated</t> U2OS cells transfected with the active LNR CC → SS 6Myc. Left upper panel shows input, and right panel streptavidin
    Streptavidin Coupled Agarose Beads, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin coupled agarose beads/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin coupled agarose beads - by Bioz Stars, 2021-05
    97/100 stars
      Buy from Supplier

    Image Search Results


    Dual-aptamer detection schematic Thrombin and PDGF-BB are represented by triangles and double-ellipses, respectively. The sensing aptamers (MB-T, MB-P) containing thrombin aptamer (Apt 1A) and PDGF-BB aptamer (Apt 2A) are each labeled with an MB tag (MB1 and MB2) and flanked by a pair of universal primers (5′p and 3′p). Capture aptamers (Bio-T and Bio-P) containing thrombin aptamer (Apt 1B) and PDGF-BB aptamer (Apt 2B) are biotinylated to be attached to streptavidin-coated magnetic beads. Oligo(dT)s are used as the spacer between the aptamer sequences and the beads. After separation, eluted sensing aptamers are amplified by multiplex PCR and quantified.

    Journal: Biosensors & bioelectronics

    Article Title: Development of a dual aptamer-based multiplex protein biosensor

    doi: 10.1016/j.bios.2010.04.034

    Figure Lengend Snippet: Dual-aptamer detection schematic Thrombin and PDGF-BB are represented by triangles and double-ellipses, respectively. The sensing aptamers (MB-T, MB-P) containing thrombin aptamer (Apt 1A) and PDGF-BB aptamer (Apt 2A) are each labeled with an MB tag (MB1 and MB2) and flanked by a pair of universal primers (5′p and 3′p). Capture aptamers (Bio-T and Bio-P) containing thrombin aptamer (Apt 1B) and PDGF-BB aptamer (Apt 2B) are biotinylated to be attached to streptavidin-coated magnetic beads. Oligo(dT)s are used as the spacer between the aptamer sequences and the beads. After separation, eluted sensing aptamers are amplified by multiplex PCR and quantified.

    Article Snippet: Binding reactions were incubated at room temperature for 30 min followed by the addition of streptavidin coated magnetic beads (Dynal/Invitrogen, Oslo, Norway) for an additional 30 min at room temperature.

    Techniques: Labeling, Magnetic Beads, Amplification, Multiplex Assay, Polymerase Chain Reaction

    Insulin-induced translocation of GLUT4 is disrupted in muscle fibers isolated from HTZ mice. ( a–c ) Freshly dissected FDB muscles from 2 month-old WT and HTZ mice were digested with collagenase. Isolated fibers were stimulated during 15 min with 0.1 µM insulin to induce GLUT4 translocation, fixed and immunolabeled with a polyclonal-GLUT4 antibody. Translocation of GLUT4 was estimated by measuring the total intensity fluorescence of GLUT4 in ROIs at the sarcolemma. ( a ) Examples of the ROIs used are drawn in white. GLUT4 was measured on both edges of the confocal image and then averaged. ( b ) Examples images of GLUT4 signal in WT and HTZ fibers at the resting (left panels) and insulin-stimulated condition (right panels). Scale bar = 20 µm. ( c ) The graph show the averaged GLUT4 signal in sarcolemma. Note that insulin-induced GLUT4 translocation is significantly reduced in HTZ myofibers compared to WT myofibers. Data are expressed as mean GLUT4 fluorescence signal ± SEM. Statistical comparisons were performed utilizing a two-tail t-test Welch corrected for parametric data. The symbols * and # denote significance with respect to WT-resting and WT-insulin-stimulated fibers, respectively. N is between 34 and 66 fibers from at least 10 different animals per genotype. ( d ) FDB muscles were dissected from WT and HTZ mice, stabilized in Tyrode solution, stimulated for 30 min with 0.1 µM insulin and then exposed to 1 mg/ml of biotin at 4 °C during 60 min. After quenching with 100 mM glycine, muscles were frozen and pulverized in liquid nitrogen, lysed and centrifuged at 14.000 g for 10 min. Supernatants were mixed with streptavidin-agarose beads overnight at 4 °C and then centrifuged at 14.000 g for 3 min. Biotinylated and non-biotinylated fractions were used to evaluate GLUT4 expression by western blot. GAPDH was used as a control that only surface proteins were labeled in biotinylated fractions. On the left are shown representative blots per each condition, on the right are plotted the percentages of GLUT4 in biotinylated fractions. Data are expressed as mean GLUT4% ± SEM. Statistical comparisons were performed utilizing a two-tail t-test Welch corrected for parametric data. The symbol * denote significance with respect to WT-muscles. N is five different animals per genotype.

    Journal: Scientific Reports

    Article Title: Dynamin-2 mutations linked to Centronuclear Myopathy impair actin-dependent trafficking in muscle cells

    doi: 10.1038/s41598-017-04418-w

    Figure Lengend Snippet: Insulin-induced translocation of GLUT4 is disrupted in muscle fibers isolated from HTZ mice. ( a–c ) Freshly dissected FDB muscles from 2 month-old WT and HTZ mice were digested with collagenase. Isolated fibers were stimulated during 15 min with 0.1 µM insulin to induce GLUT4 translocation, fixed and immunolabeled with a polyclonal-GLUT4 antibody. Translocation of GLUT4 was estimated by measuring the total intensity fluorescence of GLUT4 in ROIs at the sarcolemma. ( a ) Examples of the ROIs used are drawn in white. GLUT4 was measured on both edges of the confocal image and then averaged. ( b ) Examples images of GLUT4 signal in WT and HTZ fibers at the resting (left panels) and insulin-stimulated condition (right panels). Scale bar = 20 µm. ( c ) The graph show the averaged GLUT4 signal in sarcolemma. Note that insulin-induced GLUT4 translocation is significantly reduced in HTZ myofibers compared to WT myofibers. Data are expressed as mean GLUT4 fluorescence signal ± SEM. Statistical comparisons were performed utilizing a two-tail t-test Welch corrected for parametric data. The symbols * and # denote significance with respect to WT-resting and WT-insulin-stimulated fibers, respectively. N is between 34 and 66 fibers from at least 10 different animals per genotype. ( d ) FDB muscles were dissected from WT and HTZ mice, stabilized in Tyrode solution, stimulated for 30 min with 0.1 µM insulin and then exposed to 1 mg/ml of biotin at 4 °C during 60 min. After quenching with 100 mM glycine, muscles were frozen and pulverized in liquid nitrogen, lysed and centrifuged at 14.000 g for 10 min. Supernatants were mixed with streptavidin-agarose beads overnight at 4 °C and then centrifuged at 14.000 g for 3 min. Biotinylated and non-biotinylated fractions were used to evaluate GLUT4 expression by western blot. GAPDH was used as a control that only surface proteins were labeled in biotinylated fractions. On the left are shown representative blots per each condition, on the right are plotted the percentages of GLUT4 in biotinylated fractions. Data are expressed as mean GLUT4% ± SEM. Statistical comparisons were performed utilizing a two-tail t-test Welch corrected for parametric data. The symbol * denote significance with respect to WT-muscles. N is five different animals per genotype.

    Article Snippet: Homogenates were centrifuged at 14.000 g for 10 min and supernatants were mixed with 50 µl of streptavidin-agarose beads (Thermo Scientific #29204) overnight at 4 °C with gentle agitation and then centrifuged at 14.000 g for 3 min. Biotinylated and non-biotinylated fractions were diluted in loading buffer, subjected to SDS-PAGE and western blot to evaluate GLUT4 and GAPDH expression using specific antibodies (rabbit-polyclonal-anti GLUT4, Abcam ab654 and mouse-monoclonal-anti-GAPDH, Abcam ab9484, respectively).

    Techniques: Translocation Assay, Isolation, Mouse Assay, Immunolabeling, Fluorescence, Expressing, Western Blot, Labeling

    Notch1 S2 cleavage occurs at the cell surface. A, upper panel, Myc immunoblot of surface-biotinylated and streptavidin-precipitated U2OS cells transfected with the active LNR CC → SS 6Myc. Left upper panel shows input, and right panel streptavidin

    Journal: The Journal of Biological Chemistry

    Article Title: Metalloprotease ADAM10 Is Required for Notch1 Site 2 Cleavage *

    doi: 10.1074/jbc.M109.006775

    Figure Lengend Snippet: Notch1 S2 cleavage occurs at the cell surface. A, upper panel, Myc immunoblot of surface-biotinylated and streptavidin-precipitated U2OS cells transfected with the active LNR CC → SS 6Myc. Left upper panel shows input, and right panel streptavidin

    Article Snippet: After input, samples were taken, and streptavidin-coupled agarose beads (Pierce) were added to the supernatant and incubated for 90 min.

    Techniques: Transfection