alkaline phosphatase conjugated streptavidin  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Streptavidin Protein AP
    Description:
    Streptavidin for Western Blot ELISA
    Catalog Number:
    21323
    Price:
    None
    Applications:
    Cell Analysis|Cellular Imaging|ELISA|Protein Assays and Analysis|Protein Biology|Western Blotting
    Category:
    Antibodies Secondary Detection Reagents
    Buy from Supplier


    Structured Review

    Thermo Fisher alkaline phosphatase conjugated streptavidin
    Surface plasmon resonance analysis of biotin-tubulin binding to <t>strepavidin</t> and subsequent dimer dissociation as a result of dilution. (A) The plasmon resonance signal was increased by 954 RU during a 10-min flow of biotin-tubulin in Pi buffer with 12 mM Mg. The almost instantaneous 3000 RU signal change at the start and finish of the flow of the tubulin resulted from a difference in refractive index of the tubulin solution and the buffer. (B) Flow of tubulin- and nucleotide-free buffer resulted in a 445 RU signal decrease; the curve corresponds to a rate constant 14.72 × 10 −5 s −1 . A rate constant equal to 12.35 × 10 −5 s −1 was determined from a Guggenheim plot of the data.
    Streptavidin for Western Blot ELISA
    https://www.bioz.com/result/alkaline phosphatase conjugated streptavidin/product/Thermo Fisher
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    alkaline phosphatase conjugated streptavidin - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Dissociation of the Tubulin Dimer Is Extremely Slow, Thermodynamically Very Unfavorable, and Reversible in the Absence of an Energy Source"

    Article Title: Dissociation of the Tubulin Dimer Is Extremely Slow, Thermodynamically Very Unfavorable, and Reversible in the Absence of an Energy Source

    Journal: Molecular Biology of the Cell

    doi: 10.1091/mbc.E01-10-0089

    Surface plasmon resonance analysis of biotin-tubulin binding to strepavidin and subsequent dimer dissociation as a result of dilution. (A) The plasmon resonance signal was increased by 954 RU during a 10-min flow of biotin-tubulin in Pi buffer with 12 mM Mg. The almost instantaneous 3000 RU signal change at the start and finish of the flow of the tubulin resulted from a difference in refractive index of the tubulin solution and the buffer. (B) Flow of tubulin- and nucleotide-free buffer resulted in a 445 RU signal decrease; the curve corresponds to a rate constant 14.72 × 10 −5 s −1 . A rate constant equal to 12.35 × 10 −5 s −1 was determined from a Guggenheim plot of the data.
    Figure Legend Snippet: Surface plasmon resonance analysis of biotin-tubulin binding to strepavidin and subsequent dimer dissociation as a result of dilution. (A) The plasmon resonance signal was increased by 954 RU during a 10-min flow of biotin-tubulin in Pi buffer with 12 mM Mg. The almost instantaneous 3000 RU signal change at the start and finish of the flow of the tubulin resulted from a difference in refractive index of the tubulin solution and the buffer. (B) Flow of tubulin- and nucleotide-free buffer resulted in a 445 RU signal decrease; the curve corresponds to a rate constant 14.72 × 10 −5 s −1 . A rate constant equal to 12.35 × 10 −5 s −1 was determined from a Guggenheim plot of the data.

    Techniques Used: SPR Assay, Binding Assay, Flow Cytometry

    2) Product Images from "Disruption of Cell Polarity by Enteropathogenic Escherichia coli Enables Basolateral Membrane Proteins To Migrate Apically and To Potentiate Physiological Consequences "

    Article Title: Disruption of Cell Polarity by Enteropathogenic Escherichia coli Enables Basolateral Membrane Proteins To Migrate Apically and To Potentiate Physiological Consequences

    Journal: Infection and Immunity

    doi: 10.1128/IAI.71.12.7069-7078.2003

    (A) β 1 -Integrin is trapped on the apical surface after EDTA treatment. T84 monolayers were treated with 5 mM EDTA for 10 min to disrupt TJs and allow for redistribution of basolateral proteins to the apical surface. EDTA was then removed, and calcium was restored to reseal TJs for a 3-h recovery period. Apical surfaces of the nontreated and EDTA-treated monolayers were biotinylated, and β 1 -integrin was immunoprecipitated and detected with alkaline phosphatase-streptavidin. A significant amount of β 1 -integrin was found to be biotinylated after EDTA treatment and recovery. (B) Control monolayers and monolayers treated with EDTA were allowed to recover and were then fixed and stained for β 1 -integrin (green) and actin (red). In control monolayers β 1 -integrin is basolateral to the actin-myosin ring. However, in EDTA-treated monolayers, β 1 -integrin stains at and above this ring, indicating that is has moved to the apical pole after EDTA disruption. (C) EDTA-treated and recovered monolayers were then infected with Δ tir EPEC and stained for β 1 -integrin (green) and actin (red). Coalescing of actin staining was seen following infection, and regions of colocalized actin and β 1 -integrin can be seen. (D) Redistribution of basolateral proteins to the apical surface of cell monolayers prior to infection renders Δ tir EPEC capable of decreasing TER. T84 monolayers were treated with EDTA for 10 min, and TER was measured to confirm disruption of TJs. EDTA was then removed, and calcium was restored to reseal TJs and trap redistributed basolateral membrane proteins on the apical surface. TER was measured prior to and throughout the course of infection with wild-type EPEC or Δ tir EPEC. EPEC and Δ tir EPEC both decreased TER significantly compared to uninfected controls, with P = 4.1 × 10 −11 and P = 2.43 × 10 −5 , respectively ( n = 6). The presence of β 1 -integrin antibody throughout the course of infection blocked the Δ tir EPEC-induced decrease in TER, whereas isotype IgG had no effect (Δ tir EPEC versus Δ tir EPEC plus β 1 -integrin antibody, P = 0.002 [ n = 9]; Δ tir EPEC versus Δ tir EPEC plus IgG, P = 0.33 [ n = 5]). Error bars indicate SEMs.
    Figure Legend Snippet: (A) β 1 -Integrin is trapped on the apical surface after EDTA treatment. T84 monolayers were treated with 5 mM EDTA for 10 min to disrupt TJs and allow for redistribution of basolateral proteins to the apical surface. EDTA was then removed, and calcium was restored to reseal TJs for a 3-h recovery period. Apical surfaces of the nontreated and EDTA-treated monolayers were biotinylated, and β 1 -integrin was immunoprecipitated and detected with alkaline phosphatase-streptavidin. A significant amount of β 1 -integrin was found to be biotinylated after EDTA treatment and recovery. (B) Control monolayers and monolayers treated with EDTA were allowed to recover and were then fixed and stained for β 1 -integrin (green) and actin (red). In control monolayers β 1 -integrin is basolateral to the actin-myosin ring. However, in EDTA-treated monolayers, β 1 -integrin stains at and above this ring, indicating that is has moved to the apical pole after EDTA disruption. (C) EDTA-treated and recovered monolayers were then infected with Δ tir EPEC and stained for β 1 -integrin (green) and actin (red). Coalescing of actin staining was seen following infection, and regions of colocalized actin and β 1 -integrin can be seen. (D) Redistribution of basolateral proteins to the apical surface of cell monolayers prior to infection renders Δ tir EPEC capable of decreasing TER. T84 monolayers were treated with EDTA for 10 min, and TER was measured to confirm disruption of TJs. EDTA was then removed, and calcium was restored to reseal TJs and trap redistributed basolateral membrane proteins on the apical surface. TER was measured prior to and throughout the course of infection with wild-type EPEC or Δ tir EPEC. EPEC and Δ tir EPEC both decreased TER significantly compared to uninfected controls, with P = 4.1 × 10 −11 and P = 2.43 × 10 −5 , respectively ( n = 6). The presence of β 1 -integrin antibody throughout the course of infection blocked the Δ tir EPEC-induced decrease in TER, whereas isotype IgG had no effect (Δ tir EPEC versus Δ tir EPEC plus β 1 -integrin antibody, P = 0.002 [ n = 9]; Δ tir EPEC versus Δ tir EPEC plus IgG, P = 0.33 [ n = 5]). Error bars indicate SEMs.

    Techniques Used: Immunoprecipitation, Staining, Infection

    EPEC infection induces redistribution of basolateral membrane proteins. T84 monolayers that were either uninfected; infected with EPEC for 2, 4, or 6 h, or EDTA treated were selectively surface membrane labeled with activated biotin. Extracted proteins were immunoprecipitated with either Na + /K + ATPase or β 1 -integrin antibodies and quantified by alkaline phosphatase-streptavidin immunoblots. (A) Immunoprecipitation specific for Na + /K + ATPase in uninfected monolayers, monolayers infected for 6 h, and monolayers exposed to 4 mM EDTA for 10 min revealed a redistribution of protein from the basolateral (BL) to the apical (AP) surface following EPEC infection ( P = 0.05; n = 3) and EDTA treatment ( P = 0.004; n = 3). (B) Immunoprecipitation of β 1 -integrin was performed on uninfected monolayers; monolayers infected with EPEC for 2, 4, or 6 h; and EDTA-treated monolayers. A progressive and significant redistribution of β 1 -integrin to the apical surface was observed by 6 h postinfection ( P = 0.03; n = 4). Immunoblots are representative of those from four separate experiments. Densitometry values are expressed as mean percentages of the total (apical plus basolateral) ± SEM.
    Figure Legend Snippet: EPEC infection induces redistribution of basolateral membrane proteins. T84 monolayers that were either uninfected; infected with EPEC for 2, 4, or 6 h, or EDTA treated were selectively surface membrane labeled with activated biotin. Extracted proteins were immunoprecipitated with either Na + /K + ATPase or β 1 -integrin antibodies and quantified by alkaline phosphatase-streptavidin immunoblots. (A) Immunoprecipitation specific for Na + /K + ATPase in uninfected monolayers, monolayers infected for 6 h, and monolayers exposed to 4 mM EDTA for 10 min revealed a redistribution of protein from the basolateral (BL) to the apical (AP) surface following EPEC infection ( P = 0.05; n = 3) and EDTA treatment ( P = 0.004; n = 3). (B) Immunoprecipitation of β 1 -integrin was performed on uninfected monolayers; monolayers infected with EPEC for 2, 4, or 6 h; and EDTA-treated monolayers. A progressive and significant redistribution of β 1 -integrin to the apical surface was observed by 6 h postinfection ( P = 0.03; n = 4). Immunoblots are representative of those from four separate experiments. Densitometry values are expressed as mean percentages of the total (apical plus basolateral) ± SEM.

    Techniques Used: Infection, Labeling, Immunoprecipitation, Western Blot

    (A) The tir deletion strain CVD463 (Δ tir EPEC) has no impact on TER. Uninfected T84 monolayers and those uninfected with wild-type EPEC or EPEC Δ tir were serially assessed for TER. As shown, expression of Tir is required for the EPEC-associated decrease in TER. The data shown represent the mean ± SEM ( n = 8; P = 0.003 for EPEC versus Δ tir EPEC) from three experiments with duplicate or triplicate samples. (B) Deletion of tir also blocks redistribution of β 1 -integrin. Uninfected T84 monolayers or those infected with EPEC Δ tir or wild-type EPEC for 6 h were selectively labeled with biotin on the apical (AP) or basolateral (BL) surface, immunoprecipitated with β 1 -integrin antibody, and detected with alkaline phosphatase-streptavidin. Even after 6 h of infection, no redistribution of β 1 -integrin was detected in the monolayers infected with EPEC Δ tir (EPEC Δ tir versus wild-type EPEC after 6 h [ n = 3], P = 0.02). Densitometry data are presented as percentages of the total for the apical and basolateral surfaces. The immunoblot is representative of those from three separate experiments.
    Figure Legend Snippet: (A) The tir deletion strain CVD463 (Δ tir EPEC) has no impact on TER. Uninfected T84 monolayers and those uninfected with wild-type EPEC or EPEC Δ tir were serially assessed for TER. As shown, expression of Tir is required for the EPEC-associated decrease in TER. The data shown represent the mean ± SEM ( n = 8; P = 0.003 for EPEC versus Δ tir EPEC) from three experiments with duplicate or triplicate samples. (B) Deletion of tir also blocks redistribution of β 1 -integrin. Uninfected T84 monolayers or those infected with EPEC Δ tir or wild-type EPEC for 6 h were selectively labeled with biotin on the apical (AP) or basolateral (BL) surface, immunoprecipitated with β 1 -integrin antibody, and detected with alkaline phosphatase-streptavidin. Even after 6 h of infection, no redistribution of β 1 -integrin was detected in the monolayers infected with EPEC Δ tir (EPEC Δ tir versus wild-type EPEC after 6 h [ n = 3], P = 0.02). Densitometry data are presented as percentages of the total for the apical and basolateral surfaces. The immunoblot is representative of those from three separate experiments.

    Techniques Used: Expressing, Infection, Labeling, Immunoprecipitation

    3) Product Images from "Candidate autoantigens identified by mass spectrometry in early rheumatoid arthritis are chaperones and citrullinated glycolytic enzymes"

    Article Title: Candidate autoantigens identified by mass spectrometry in early rheumatoid arthritis are chaperones and citrullinated glycolytic enzymes

    Journal: Arthritis Research & Therapy

    doi: 10.1186/ar2644

    Detection of autoantibodies in rheumatoid arthritis patient sera. Autoantibodies in rheumatoid arthritis (RA) patient sera were detected by western blot analysis using HL-60 cell extract as the substrate. (a) Example of one-dimensional gel electrophoresis western blot analysis with Imagemaster totalLab software to determine the molecular weights (m.w.) of different bands using an internal standard (is1 and is2) that correspond to 120-kDa and 80-kDa proteins revealed by alkaline phosphatase-conjugated streptavidin. These bands were used for standardization between the different membranes. (b) Virtual blot of the 110 RA patient sera. The m.w. of the bands are indicated on the right-hand side of the figure. Each vertical lane corresponds to different RA patient sera.
    Figure Legend Snippet: Detection of autoantibodies in rheumatoid arthritis patient sera. Autoantibodies in rheumatoid arthritis (RA) patient sera were detected by western blot analysis using HL-60 cell extract as the substrate. (a) Example of one-dimensional gel electrophoresis western blot analysis with Imagemaster totalLab software to determine the molecular weights (m.w.) of different bands using an internal standard (is1 and is2) that correspond to 120-kDa and 80-kDa proteins revealed by alkaline phosphatase-conjugated streptavidin. These bands were used for standardization between the different membranes. (b) Virtual blot of the 110 RA patient sera. The m.w. of the bands are indicated on the right-hand side of the figure. Each vertical lane corresponds to different RA patient sera.

    Techniques Used: Western Blot, Nucleic Acid Electrophoresis, Software

    Detection of citrullinated proteins on two-dimensional gel electrophoresis HL-60 protein maps. Citrullinated proteins were detected on two-dimensional gel electrophoresis HL-60 protein maps (a) before and (b) after treatment with peptidyl-arginine deiminase. The membranes were incubated with rabbit anti-citrulline antibodies, washed and then incubated with biotinylated-goat anti-rabbit antibodies followed by IRDye 800-conjugated streptavidin, and were visualized using the Odyssey™ Infrared Imaging system. FUSE-BP, far-upstream element-binding protein; HnRNP A2/B1, heterogeneous nuclear ribonucleoprotein A2/B1; HSP60, 60 kDa heat shock protein; m.w., molecular weight; PGK, phosphoglycerate kinase 1.
    Figure Legend Snippet: Detection of citrullinated proteins on two-dimensional gel electrophoresis HL-60 protein maps. Citrullinated proteins were detected on two-dimensional gel electrophoresis HL-60 protein maps (a) before and (b) after treatment with peptidyl-arginine deiminase. The membranes were incubated with rabbit anti-citrulline antibodies, washed and then incubated with biotinylated-goat anti-rabbit antibodies followed by IRDye 800-conjugated streptavidin, and were visualized using the Odyssey™ Infrared Imaging system. FUSE-BP, far-upstream element-binding protein; HnRNP A2/B1, heterogeneous nuclear ribonucleoprotein A2/B1; HSP60, 60 kDa heat shock protein; m.w., molecular weight; PGK, phosphoglycerate kinase 1.

    Techniques Used: Two-Dimensional Gel Electrophoresis, Electrophoresis, Incubation, Imaging, Binding Assay, Molecular Weight

    4) Product Images from "In vivo alteration of humoral responses to HIV-1 envelope glycoprotein gp120 by antibodies to the CD4-binding site of gp120"

    Article Title: In vivo alteration of humoral responses to HIV-1 envelope glycoprotein gp120 by antibodies to the CD4-binding site of gp120

    Journal: Virology

    doi: 10.1016/j.virol.2007.10.044

    Reactivity of mAbs to V3, C1, and C5 with gp120/mAb complexes. (A) gp120 or gp120/mAb complexes were captured on the ELISA plates by polyclonal anti-C5 Abs and reacted with biotinylated mAbs specific for V3 (694/98D and 447/52D) or C1 (EH21). Alternatively, gp120 or the complexes were directly coated onto ELISA plates and reacted with biotinylated mAbs specific for C5 (1331A). The gp120/mAb complexes were prepared with 1 μg/ml of recombinant gp120 LAI and 2 μg/ml of mAbs to the CD4bs (654D) or C2 (1006-30D) and serially diluted by 2 fold. Gp120 mixed with a control mAb (1418) was also included as a control. The x-axis shows the gp120 concentrations in each of the gp120/mAb mixtures. Relative binding of the biotinylated mAbs was determined using alkaline phosphatase-conjugated streptavidin. Means and standard deviations were calculated from duplicate wells. Data from one of five repeated experiments are shown. *, p
    Figure Legend Snippet: Reactivity of mAbs to V3, C1, and C5 with gp120/mAb complexes. (A) gp120 or gp120/mAb complexes were captured on the ELISA plates by polyclonal anti-C5 Abs and reacted with biotinylated mAbs specific for V3 (694/98D and 447/52D) or C1 (EH21). Alternatively, gp120 or the complexes were directly coated onto ELISA plates and reacted with biotinylated mAbs specific for C5 (1331A). The gp120/mAb complexes were prepared with 1 μg/ml of recombinant gp120 LAI and 2 μg/ml of mAbs to the CD4bs (654D) or C2 (1006-30D) and serially diluted by 2 fold. Gp120 mixed with a control mAb (1418) was also included as a control. The x-axis shows the gp120 concentrations in each of the gp120/mAb mixtures. Relative binding of the biotinylated mAbs was determined using alkaline phosphatase-conjugated streptavidin. Means and standard deviations were calculated from duplicate wells. Data from one of five repeated experiments are shown. *, p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Recombinant, Binding Assay

    5) Product Images from "Nitrosative stress in human skeletal muscle attenuated by exercise countermeasure after chronic disuse"

    Article Title: Nitrosative stress in human skeletal muscle attenuated by exercise countermeasure after chronic disuse

    Journal: Redox Biology

    doi: 10.1016/j.redox.2013.10.006

    BST detection of sarcoplasmic reticulum SNO-SERCA1/–SERCA2 in human skeletal muscle SOL and VL biopsies from bed rest groups. A. Upper panel , Immunoblot of SERCA1 in BST streptavidin column eluate of SOL; lower panel , percent changes SNO-Cys-SERCA1 and SNO-Cys-SERCA2 of End vs. Pre bed rest SOL (Pre values are set up as zero baselines). An increase of SNO-SERCA1 in CTR (330%, p ≤0.01, n =9), RE (180%, p ≤0.01, n =7), and RVE (75%, p ≤0.01, n =7) group was detected (*). No changes were observed for SNO-SERCA2 proteins. B. Upper panel , Immunoblot of SERCA1 of streptavidin column eluate of VL; lower panel , percent changes SNO-Cys-SERCA1 and SNO-Cys-SERCA2 of End vs. Pre bed rest VL (Pre values are set up as zero baselines). An increase of SERCA1 (220%, p ≤0.01, n =9) was present in CTR End subjects, while a decrease was present in RE (-41%, p ≤0.01, n =7) and RVE (-40%, p ≤0.01, n =7) subjects (*). No changes were observed for SNO-SERCA2 proteins.
    Figure Legend Snippet: BST detection of sarcoplasmic reticulum SNO-SERCA1/–SERCA2 in human skeletal muscle SOL and VL biopsies from bed rest groups. A. Upper panel , Immunoblot of SERCA1 in BST streptavidin column eluate of SOL; lower panel , percent changes SNO-Cys-SERCA1 and SNO-Cys-SERCA2 of End vs. Pre bed rest SOL (Pre values are set up as zero baselines). An increase of SNO-SERCA1 in CTR (330%, p ≤0.01, n =9), RE (180%, p ≤0.01, n =7), and RVE (75%, p ≤0.01, n =7) group was detected (*). No changes were observed for SNO-SERCA2 proteins. B. Upper panel , Immunoblot of SERCA1 of streptavidin column eluate of VL; lower panel , percent changes SNO-Cys-SERCA1 and SNO-Cys-SERCA2 of End vs. Pre bed rest VL (Pre values are set up as zero baselines). An increase of SERCA1 (220%, p ≤0.01, n =9) was present in CTR End subjects, while a decrease was present in RE (-41%, p ≤0.01, n =7) and RVE (-40%, p ≤0.01, n =7) subjects (*). No changes were observed for SNO-SERCA2 proteins.

    Techniques Used:

    BST detection of sarcolemmal and sarcoplasmatic reticulum functional channel proteins in human skeletal SOL and VL from bed rest subjects. A. Upper and lower panel , DHPR1 α immunoblot of BST streptavidin column eluate of SOL and VL. A moderate presence of DHPR1 α proteins was present in all groups (CTR n =9; RE n =7; RVE n =7). B. Upper and lower panel , IP 3 R1 immunoblot of BST streptavidin column eluate of SOL and VL. A moderate presence of IP 3 R1 proteins was present in all groups (Pre). No changes were present in both samples after bed rest (End) (CTR n =9; RE n =7; RVE n =7). C. Left and right panel , TRPC1 immunoblot of BST streptavidin column eluate of SOL and VL (CTR n =9; RE n =7; RVE n =7); (Contr.=positive control of total proteins muscle lysates). No SNO-TRPC1 protein signals were detectable in either group before and after bed rest.
    Figure Legend Snippet: BST detection of sarcolemmal and sarcoplasmatic reticulum functional channel proteins in human skeletal SOL and VL from bed rest subjects. A. Upper and lower panel , DHPR1 α immunoblot of BST streptavidin column eluate of SOL and VL. A moderate presence of DHPR1 α proteins was present in all groups (CTR n =9; RE n =7; RVE n =7). B. Upper and lower panel , IP 3 R1 immunoblot of BST streptavidin column eluate of SOL and VL. A moderate presence of IP 3 R1 proteins was present in all groups (Pre). No changes were present in both samples after bed rest (End) (CTR n =9; RE n =7; RVE n =7). C. Left and right panel , TRPC1 immunoblot of BST streptavidin column eluate of SOL and VL (CTR n =9; RE n =7; RVE n =7); (Contr.=positive control of total proteins muscle lysates). No SNO-TRPC1 protein signals were detectable in either group before and after bed rest.

    Techniques Used: Functional Assay, Positive Control

    BST detection of SNO-RyR1 in human skeletal muscle SOL and VL biopsies. A. Upper panel , Immunoblot of BST streptavidin column eluate of SOL for RyR1. A. Lower panel , Immunoblot of BST streptavidin column eluate of VL for RyR1. Increased RyR1 immunoreactive band was detected in CTR End-only samples. B. Percent changes biotin-labeled RyR1 proteins from Pre and End bed rest biopsies. Bed rest without exercise promoted S -nitrosylation of RyR1 in disused SOL (CTR n =9, 18.6%, p ≤0.01) and VL (CTR n =9, 17.2%, p ≤0.01) as counteracted by both exercise countermeasures. (* significance).
    Figure Legend Snippet: BST detection of SNO-RyR1 in human skeletal muscle SOL and VL biopsies. A. Upper panel , Immunoblot of BST streptavidin column eluate of SOL for RyR1. A. Lower panel , Immunoblot of BST streptavidin column eluate of VL for RyR1. Increased RyR1 immunoreactive band was detected in CTR End-only samples. B. Percent changes biotin-labeled RyR1 proteins from Pre and End bed rest biopsies. Bed rest without exercise promoted S -nitrosylation of RyR1 in disused SOL (CTR n =9, 18.6%, p ≤0.01) and VL (CTR n =9, 17.2%, p ≤0.01) as counteracted by both exercise countermeasures. (* significance).

    Techniques Used: Labeling

    SNO-protein assay in human skeletal muscle biopsies. A. Dot blots analysis (left panel) biotin-labeled proteins (reflecting SNO-proteins) in soleus (SOL) of End vs. Pre bed rest subjects (1,2,3: triplicate), Control BST was obtained by omitting biotin-HPDP during the protocol; right panel, dot blot densitometry analysis. B. (left panel) Dot blot analysis of biotin-labeled proteins in vastus lateralis (VL) of End vs. Pre bed rest subjects (1,2,3: triplicate), Control BST was obtained by omitting biotin-HPDP during the protocol; right panel, dot blot densitometry analysis. A significant increase of biotin incorporation is observed in CTR group ( n =9) in both muscles (SOL, Pre 86.51±2.9, End 95.64±6.5; VL Pre 192.94±3.07, End 222.59±6.6). In the RE group ( n =7) a significant increase was in VL only (Pre 123.07±4.7, End 145.6±5.56) but not in SOL after bed rest. RVE ( n =7) only significantly decreased SNO-protein levels in both SOL (Pre 75.3±0.75, End 62.25±1.7) and VL (Pre 144.52±3.8, End 122.32±4.1). C. NOS1 WB analysis of BST streptavidin column eluate vs. normal muscle lysates (Muscle lysate). In both SOL and VL, NOS1 immunoreactive bands with higher relative mobility are detectable (BST/Eluate) vs. a predicted NOS1 immunoreactive band obtained from total muscle lysates used as positive control.
    Figure Legend Snippet: SNO-protein assay in human skeletal muscle biopsies. A. Dot blots analysis (left panel) biotin-labeled proteins (reflecting SNO-proteins) in soleus (SOL) of End vs. Pre bed rest subjects (1,2,3: triplicate), Control BST was obtained by omitting biotin-HPDP during the protocol; right panel, dot blot densitometry analysis. B. (left panel) Dot blot analysis of biotin-labeled proteins in vastus lateralis (VL) of End vs. Pre bed rest subjects (1,2,3: triplicate), Control BST was obtained by omitting biotin-HPDP during the protocol; right panel, dot blot densitometry analysis. A significant increase of biotin incorporation is observed in CTR group ( n =9) in both muscles (SOL, Pre 86.51±2.9, End 95.64±6.5; VL Pre 192.94±3.07, End 222.59±6.6). In the RE group ( n =7) a significant increase was in VL only (Pre 123.07±4.7, End 145.6±5.56) but not in SOL after bed rest. RVE ( n =7) only significantly decreased SNO-protein levels in both SOL (Pre 75.3±0.75, End 62.25±1.7) and VL (Pre 144.52±3.8, End 122.32±4.1). C. NOS1 WB analysis of BST streptavidin column eluate vs. normal muscle lysates (Muscle lysate). In both SOL and VL, NOS1 immunoreactive bands with higher relative mobility are detectable (BST/Eluate) vs. a predicted NOS1 immunoreactive band obtained from total muscle lysates used as positive control.

    Techniques Used: Labeling, Dot Blot, Western Blot, Positive Control

    BST-related SNO-NOS1 protein assay of human SOL and VL muscle biopsies before and after bed rest. A. Upper panel , NOS1 WB analysis of SOL lysates normalized to alpha tubulin. A. Middle panel, NOS1 WB analysis of BST-streptavidin column eluate in SOL lysates. NOS1 protein immunoreactive bands reflecting biotin-labeled NOS1 proteins were present in all eluate samples showing the presence of S-nitrosylated NOS1 proteins in human SOL. A. Lower panel , graph representing percent changes of NOS1 (white columns, CTR ( n =9), −32%, p ≤0.01; RE ( n =7), 12.7%, p ≤0.05; RVE ( n =7), 31.2%, p ≤0.01) and of SNO-NOS1 (black columns, CTR ( n =9), −40%, p ≤0.01; RE ( n =7), 42.5%, p ≤0.01; RVE ( n =7), 60.6%, p ≤0.01) proteins in SOL of End vs. Pre bed rest biopsies (Pre values are set up as zero baselines). B. Upper panel , NOS1 WB analysis of VL lysates normalized to alpha tubulin. B. Middle panel, NOS1 WB analysis of BST-streptavidin column eluate in VL lysates. NOS1 protein immunoreactive bands reflecting biotin-labeled NOS1 proteins were present in all eluate samples showing the presence of S-nitrosylated NOS1 proteins in human VL. B. Lower panel , graph representing percent changes of NOS1 (white columns, CTR ( n =9), −9.84%, p ≤0.05; RE ( n =7), 16.2%, p ≤0.01; RVE ( n =7), 9.5%, p ≤0.05) and of SNO-NOS1 (black columns, CTR ( n =9), −12%, p ≤0.05; RE ( n =7), 10.45%, p ≤0.05; RVE ( n =7), 15.65%, p ≤0.01) proteins in VL lysates of End vs. Pre bed rest samples (Pre values are set up as zero baselines).
    Figure Legend Snippet: BST-related SNO-NOS1 protein assay of human SOL and VL muscle biopsies before and after bed rest. A. Upper panel , NOS1 WB analysis of SOL lysates normalized to alpha tubulin. A. Middle panel, NOS1 WB analysis of BST-streptavidin column eluate in SOL lysates. NOS1 protein immunoreactive bands reflecting biotin-labeled NOS1 proteins were present in all eluate samples showing the presence of S-nitrosylated NOS1 proteins in human SOL. A. Lower panel , graph representing percent changes of NOS1 (white columns, CTR ( n =9), −32%, p ≤0.01; RE ( n =7), 12.7%, p ≤0.05; RVE ( n =7), 31.2%, p ≤0.01) and of SNO-NOS1 (black columns, CTR ( n =9), −40%, p ≤0.01; RE ( n =7), 42.5%, p ≤0.01; RVE ( n =7), 60.6%, p ≤0.01) proteins in SOL of End vs. Pre bed rest biopsies (Pre values are set up as zero baselines). B. Upper panel , NOS1 WB analysis of VL lysates normalized to alpha tubulin. B. Middle panel, NOS1 WB analysis of BST-streptavidin column eluate in VL lysates. NOS1 protein immunoreactive bands reflecting biotin-labeled NOS1 proteins were present in all eluate samples showing the presence of S-nitrosylated NOS1 proteins in human VL. B. Lower panel , graph representing percent changes of NOS1 (white columns, CTR ( n =9), −9.84%, p ≤0.05; RE ( n =7), 16.2%, p ≤0.01; RVE ( n =7), 9.5%, p ≤0.05) and of SNO-NOS1 (black columns, CTR ( n =9), −12%, p ≤0.05; RE ( n =7), 10.45%, p ≤0.05; RVE ( n =7), 15.65%, p ≤0.01) proteins in VL lysates of End vs. Pre bed rest samples (Pre values are set up as zero baselines).

    Techniques Used: Western Blot, Labeling

    BST detection of SNO-myosin heavy chain (MyHC) proteins in bed rest muscle biopsies. A. Immunoblotted BST streptavidin column eluate of SOL investigated for the presence of fast- ( upper panel ) and slow-type ( lower panel ) MyHC. Significant decrease in slow-type (−22%, p ≤0.05) and increase in fast-type (138%, p ≤0.01) MyHC was present in CTR ( n =9) End bed rest biopsies. In the RE group ( n =7), an increase of slow-type (51%, p ≤0.01) and a decrease of fast-type (−99%, p ≤0.01) MyHC was seen. In the RVE ( n =4) group, both slow-type (−99%, p ≤0.01) and fast-type (−16%, p ≤0.05) MyHCs levels were equally decreased. B. Slow- and fast-type MyHC (s/fMyHC) immunoblot analysis of BST streptavidin column eluate of VL. Significant increase of slow-type (13%, p ≤0.05, lower panel ) and fast-type (300%, p ≤0.01) MyHCs were present in CTR ( n =9) End bed rest biopsies. In the RE group ( n =7), a decrease of slow-type (−92%, p ≤0.01) and fast-type (−15%, p ≤0.05) MyHC was present. In the RVE group ( n =7), both slow- (−95%, p ≤0.01) and fast-type (−80%, p ≤0.01) MyHC levels were equally decreased.
    Figure Legend Snippet: BST detection of SNO-myosin heavy chain (MyHC) proteins in bed rest muscle biopsies. A. Immunoblotted BST streptavidin column eluate of SOL investigated for the presence of fast- ( upper panel ) and slow-type ( lower panel ) MyHC. Significant decrease in slow-type (−22%, p ≤0.05) and increase in fast-type (138%, p ≤0.01) MyHC was present in CTR ( n =9) End bed rest biopsies. In the RE group ( n =7), an increase of slow-type (51%, p ≤0.01) and a decrease of fast-type (−99%, p ≤0.01) MyHC was seen. In the RVE ( n =4) group, both slow-type (−99%, p ≤0.01) and fast-type (−16%, p ≤0.05) MyHCs levels were equally decreased. B. Slow- and fast-type MyHC (s/fMyHC) immunoblot analysis of BST streptavidin column eluate of VL. Significant increase of slow-type (13%, p ≤0.05, lower panel ) and fast-type (300%, p ≤0.01) MyHCs were present in CTR ( n =9) End bed rest biopsies. In the RE group ( n =7), a decrease of slow-type (−92%, p ≤0.01) and fast-type (−15%, p ≤0.05) MyHC was present. In the RVE group ( n =7), both slow- (−95%, p ≤0.01) and fast-type (−80%, p ≤0.01) MyHC levels were equally decreased.

    Techniques Used:

    BST detection of sarcoplasmic membrane SNO-PMCA1 in human skeletal muscle SOL and VL from bed rest groups. A. Upper panel , PMCA1 immunoblot BST streptavidin column eluate of SOL (CTR n =9; RE n =7; RVE n =7); a faint PMCA1 immunoreactive band was present in all groups. A. Lower panel , PMCA1 immunoblot of streptavidin column eluate of VL (CTR n =9; RE n =7; RVE n =7); a faint PMCA1 immunoreactive band was present in all groups. Control=positive control with human endothelial cell lysates (BD Biosciences). B. Percent change SNO-PMCA1 in SOL and VL of all groups. No changes in SNO-PMCA1 were found SOL (CTR 9.4%, p > 0.05; RE SOL 7.5%, p > 0.05, RVE 1.78%, p > 0.05) while changes were present in VL (CTR 45.55%, p ≤0.01; RE −21.35%, p ≤0.01; RVE −19.07%, p ≤0.01) after bed rest (* significance).
    Figure Legend Snippet: BST detection of sarcoplasmic membrane SNO-PMCA1 in human skeletal muscle SOL and VL from bed rest groups. A. Upper panel , PMCA1 immunoblot BST streptavidin column eluate of SOL (CTR n =9; RE n =7; RVE n =7); a faint PMCA1 immunoreactive band was present in all groups. A. Lower panel , PMCA1 immunoblot of streptavidin column eluate of VL (CTR n =9; RE n =7; RVE n =7); a faint PMCA1 immunoreactive band was present in all groups. Control=positive control with human endothelial cell lysates (BD Biosciences). B. Percent change SNO-PMCA1 in SOL and VL of all groups. No changes in SNO-PMCA1 were found SOL (CTR 9.4%, p > 0.05; RE SOL 7.5%, p > 0.05, RVE 1.78%, p > 0.05) while changes were present in VL (CTR 45.55%, p ≤0.01; RE −21.35%, p ≤0.01; RVE −19.07%, p ≤0.01) after bed rest (* significance).

    Techniques Used: Positive Control

    6) Product Images from "Nano-guided cell networks as conveyors of molecular communication"

    Article Title: Nano-guided cell networks as conveyors of molecular communication

    Journal: Nature Communications

    doi: 10.1038/ncomms9500

    Cells equipped with magnetic nanoparticles (mNPs) via streptavidin-mediated interaction with surface-expressed proteins. ( a ) Cell surface binding of streptavidin-conjugated magnetic nanoparticles occurs via surface-anchored streptavidin-binding peptide (SBP). The fusion of T7-expressed SBP-fluorescent protein (FP)-AIDAc enables the cell surface accessibility. ( b ) Scanning electron micrograph of an E. coli cell with surface-bound particles. ( c ) Element map of carbon (red) and iron (green) through energy-dispersive spectroscopy.
    Figure Legend Snippet: Cells equipped with magnetic nanoparticles (mNPs) via streptavidin-mediated interaction with surface-expressed proteins. ( a ) Cell surface binding of streptavidin-conjugated magnetic nanoparticles occurs via surface-anchored streptavidin-binding peptide (SBP). The fusion of T7-expressed SBP-fluorescent protein (FP)-AIDAc enables the cell surface accessibility. ( b ) Scanning electron micrograph of an E. coli cell with surface-bound particles. ( c ) Element map of carbon (red) and iron (green) through energy-dispersive spectroscopy.

    Techniques Used: Binding Assay, Spectroscopy

    Cells express functional, interchangeable protein components indicating both fluorescence and ability for streptavidin-linked surface coupling. ( a ) A T7 cassette was used to express chimeric proteins consisting of a membrane autotransporter domain (AIDAc), one of several fluorescent proteins and a streptavidin-binding peptide (SBP). Fluorophore-tagged streptavidin (SA) was used to bind SBP. ( b ) Of cells expressing fluorescent proteins (FP), those also marked by SBP coupling are represented as a ‘colocalized fraction ( f c ),' plotted with image analysis-based s.d. of at least five replicates. The asterisk ‘*' denotes f c that +SBP–eGFP and+SBP–mCherry are statistically equivalent ( f c ∼0.7) by t -test and greater than +SBP–Venus cells. ( c ) Composite images show cell fluorescence (Column I) from the fluorescent protein (FP); labelled streptavidin using orthogonal filter sets (Column II); and an overlay of both (Column III). Arrows indicate representative cells with strong colocalization. Plotted in Column IV are the fluorescence mean grey values ( y -axis) from a representative horizontal slice of the composite image ( x -axis). Vertical bars displayed between Columns III and IV identify the position of each analysed slice. Arrows indicate peaks that match the highlighted cells in Column III. f c values are noted. Fluorophores with non-overlapping spectra were paired. Row 1, Venus expression (yellow-green) was paired with Dylight405-labelled SA (blue). Row 2, eGFP expression (green) was paired with Alexafluor594-labeled SA (red). Row 3, mCherry expression (red) was paired with Alexafluor488-labeled SA (green). Scale bar in lower left, 50 μm.
    Figure Legend Snippet: Cells express functional, interchangeable protein components indicating both fluorescence and ability for streptavidin-linked surface coupling. ( a ) A T7 cassette was used to express chimeric proteins consisting of a membrane autotransporter domain (AIDAc), one of several fluorescent proteins and a streptavidin-binding peptide (SBP). Fluorophore-tagged streptavidin (SA) was used to bind SBP. ( b ) Of cells expressing fluorescent proteins (FP), those also marked by SBP coupling are represented as a ‘colocalized fraction ( f c ),' plotted with image analysis-based s.d. of at least five replicates. The asterisk ‘*' denotes f c that +SBP–eGFP and+SBP–mCherry are statistically equivalent ( f c ∼0.7) by t -test and greater than +SBP–Venus cells. ( c ) Composite images show cell fluorescence (Column I) from the fluorescent protein (FP); labelled streptavidin using orthogonal filter sets (Column II); and an overlay of both (Column III). Arrows indicate representative cells with strong colocalization. Plotted in Column IV are the fluorescence mean grey values ( y -axis) from a representative horizontal slice of the composite image ( x -axis). Vertical bars displayed between Columns III and IV identify the position of each analysed slice. Arrows indicate peaks that match the highlighted cells in Column III. f c values are noted. Fluorophores with non-overlapping spectra were paired. Row 1, Venus expression (yellow-green) was paired with Dylight405-labelled SA (blue). Row 2, eGFP expression (green) was paired with Alexafluor594-labeled SA (red). Row 3, mCherry expression (red) was paired with Alexafluor488-labeled SA (green). Scale bar in lower left, 50 μm.

    Techniques Used: Functional Assay, Fluorescence, Binding Assay, Expressing, Labeling

    Affinity-based probing for functional analysis of AI-2-induced protein expression. ( a ) 64–82 kDa region of western blot for pelleted (P) and supernatant (S) protein fractions isolated from Type A and B cells. Alkaline phosphatase-conjugated streptavidin was used to target AIDAc–Venus–SBP at expression timepoints. Arrows indicate the expected position of the full fusion protein. ( b ) Immunostaining for assessment of the fluorescent protein surface accessibility. The external surfaces of cells expressing AIDAc–eGFP–SBP were probed with an anti–eGFP and Alexafluor594-labelled antibody pair. A representative overlaid fluorescence and phase contrast image is shown along with fluorescence images of the green (G) and red (R) filters for the boxed-in region. Scale bar, 2 μM.
    Figure Legend Snippet: Affinity-based probing for functional analysis of AI-2-induced protein expression. ( a ) 64–82 kDa region of western blot for pelleted (P) and supernatant (S) protein fractions isolated from Type A and B cells. Alkaline phosphatase-conjugated streptavidin was used to target AIDAc–Venus–SBP at expression timepoints. Arrows indicate the expected position of the full fusion protein. ( b ) Immunostaining for assessment of the fluorescent protein surface accessibility. The external surfaces of cells expressing AIDAc–eGFP–SBP were probed with an anti–eGFP and Alexafluor594-labelled antibody pair. A representative overlaid fluorescence and phase contrast image is shown along with fluorescence images of the green (G) and red (R) filters for the boxed-in region. Scale bar, 2 μM.

    Techniques Used: Functional Assay, Expressing, Western Blot, Isolation, Immunostaining, Fluorescence

    7) Product Images from "Functional Consequences of Complementarity-determining Region Deactivation in a Multifunctional Anti-nucleic Acid Antibody *"

    Article Title: Functional Consequences of Complementarity-determining Region Deactivation in a Multifunctional Anti-nucleic Acid Antibody *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M113.508499

    Biochemical properties of 3D8 CDR-derived peptides. A and B , ELISA for DNA and heparin binding activity. Wells coated with 10 μg/ml plasmid DNA ( A ) or 1 μg/ml heparin ( B ) were incubated with biotin-labeled peptides. The bound peptides were detected using alkaline phosphatase-conjugated streptavidin. Data represent the mean ± S.D. of triplicate wells and are representative of two independent experiments. C , FRET-based DNA cleavage assay. scFvs (1 μ m ) or a mixture of scFv (1 μ m ) and heparin (10 μg/ml) were incubated with a double-labeled single-stranded DNA substrate (500 n m ) as described under “Experimental Procedures.” The fluorescence intensity was then measured in real time over 6 h (at 5-min intervals). RFU , relative fluorescence units. Data are representative of three independent experiments. D and E , cell-penetrating activity of the peptides. HeLa cells were incubated with FITC-labeled peptides (5 μ m ) for 6 h at 37 °C and then analyzed by flow cytometry ( D ) and confocal microscopy ( E ). Nuclei were stained with Hoechst 33342 ( blue ). Scale bar = 10 μm. Data are representative of three independent experiments.
    Figure Legend Snippet: Biochemical properties of 3D8 CDR-derived peptides. A and B , ELISA for DNA and heparin binding activity. Wells coated with 10 μg/ml plasmid DNA ( A ) or 1 μg/ml heparin ( B ) were incubated with biotin-labeled peptides. The bound peptides were detected using alkaline phosphatase-conjugated streptavidin. Data represent the mean ± S.D. of triplicate wells and are representative of two independent experiments. C , FRET-based DNA cleavage assay. scFvs (1 μ m ) or a mixture of scFv (1 μ m ) and heparin (10 μg/ml) were incubated with a double-labeled single-stranded DNA substrate (500 n m ) as described under “Experimental Procedures.” The fluorescence intensity was then measured in real time over 6 h (at 5-min intervals). RFU , relative fluorescence units. Data are representative of three independent experiments. D and E , cell-penetrating activity of the peptides. HeLa cells were incubated with FITC-labeled peptides (5 μ m ) for 6 h at 37 °C and then analyzed by flow cytometry ( D ) and confocal microscopy ( E ). Nuclei were stained with Hoechst 33342 ( blue ). Scale bar = 10 μm. Data are representative of three independent experiments.

    Techniques Used: Derivative Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Activity Assay, Plasmid Preparation, Incubation, Labeling, DNA Cleavage Assay, Fluorescence, Flow Cytometry, Cytometry, Confocal Microscopy, Staining

    8) Product Images from "Blocking transcription of the human rhodopsin gene by triplex-mediated DNA photocrosslinking"

    Article Title: Blocking transcription of the human rhodopsin gene by triplex-mediated DNA photocrosslinking

    Journal: Nucleic Acids Research

    doi:

    Formation of TFO-targeted crosslinks within living cells. ( A ) FACS results from control cells and cells treated with pSRG and psoralen–TFO2 and/or psoralen–TFO9 prior to UVA irradiation of the washed cells. Grey bars represent samples that did not receive any UVA irradiation after transfection, whereas black bars show samples that were irradiated 1 h after transfection. Three independent experiments were carried out and the results plotted are the means, normalized to the control (transfection with plasmid but no TFO), for each condition, with error bars indicating the standard error. ( B ) Southern blot of plasmid DNA extracted from cells after treatment with pSRG or pSRG-M2 and psoralen–TFO2–biotin prior to UVA irradiation of the washed cells (lanes 2 and 3) and from cells transfected with the TFO already crosslinked to pSRG (lane 1). (Top) The result of probing with alkaline phosphatase-conjugated streptavidin followed by recording of chemiluminescence for detection of the biotin tag. (Bottom) The result of probing with a 32 P-labeled probe specific for the plasmid fragment containing the triplex site, as a control for equal loading and transfer, and to verify that the photoadduct corresponds to the intended fragment. Arrows show the migration position corresponding to 2.2 kb.
    Figure Legend Snippet: Formation of TFO-targeted crosslinks within living cells. ( A ) FACS results from control cells and cells treated with pSRG and psoralen–TFO2 and/or psoralen–TFO9 prior to UVA irradiation of the washed cells. Grey bars represent samples that did not receive any UVA irradiation after transfection, whereas black bars show samples that were irradiated 1 h after transfection. Three independent experiments were carried out and the results plotted are the means, normalized to the control (transfection with plasmid but no TFO), for each condition, with error bars indicating the standard error. ( B ) Southern blot of plasmid DNA extracted from cells after treatment with pSRG or pSRG-M2 and psoralen–TFO2–biotin prior to UVA irradiation of the washed cells (lanes 2 and 3) and from cells transfected with the TFO already crosslinked to pSRG (lane 1). (Top) The result of probing with alkaline phosphatase-conjugated streptavidin followed by recording of chemiluminescence for detection of the biotin tag. (Bottom) The result of probing with a 32 P-labeled probe specific for the plasmid fragment containing the triplex site, as a control for equal loading and transfer, and to verify that the photoadduct corresponds to the intended fragment. Arrows show the migration position corresponding to 2.2 kb.

    Techniques Used: FACS, Irradiation, Transfection, Plasmid Preparation, Southern Blot, Labeling, Migration

    9) Product Images from "3?-End Formation of Baculovirus Late RNAs"

    Article Title: 3?-End Formation of Baculovirus Late RNAs

    Journal: Journal of Virology

    doi:

    Mapping of terminated transcripts. (A) Schematic diagram of Polh/CFS-T with the relative positions of the 5′ and 3′ probes indicated. (B) Transcription reactions. RNA transcripts were resolved on a 6% polyacrylamide–8 M urea gel, transferred to a nylon membrane, and detected by exposure to X-ray film. Lanes: M, φX174/ Hin fI marker; 1 and 3, Polh/CFS as template; 2 and 4, Polh/CFS-T as template. (C) Northern blot analysis. The nylon membrane was hybridized with biotin-labeled probes and detected using alkaline phosphatase-conjugated streptavidin and a chromogenic substrate. Sizes are indicated in nucleotides.
    Figure Legend Snippet: Mapping of terminated transcripts. (A) Schematic diagram of Polh/CFS-T with the relative positions of the 5′ and 3′ probes indicated. (B) Transcription reactions. RNA transcripts were resolved on a 6% polyacrylamide–8 M urea gel, transferred to a nylon membrane, and detected by exposure to X-ray film. Lanes: M, φX174/ Hin fI marker; 1 and 3, Polh/CFS as template; 2 and 4, Polh/CFS-T as template. (C) Northern blot analysis. The nylon membrane was hybridized with biotin-labeled probes and detected using alkaline phosphatase-conjugated streptavidin and a chromogenic substrate. Sizes are indicated in nucleotides.

    Techniques Used: Marker, Northern Blot, Labeling

    10) Product Images from "Decorin Binding by DbpA and B of Borrelia garinii, Borrelia afzelii, and Borrelia burgdorferi Sensu Stricto"

    Article Title: Decorin Binding by DbpA and B of Borrelia garinii, Borrelia afzelii, and Borrelia burgdorferi Sensu Stricto

    Journal: The Journal of Infectious Diseases

    doi: 10.1093/infdis/jir207

    A , Binding of decorin to recombinant Dbps in a Western blot assay. The upper panel represents SimplyBlue staining of the SDS-PAGE gel. The lower panel shows the adhesion of biotinylated decorin (1 μg/mL) to Dbps. B , Binding of decorin to recombinant Dbps in a microtiter plate assay. Recombinant Dbps (10 μg/mL) were attached on microtiter plates, and unspecific binding was blocked with BSA. The binding of biotinylated decorin (1 μg/mL) was detected with alkaline-phosphatase streptavidin and p-NPP-Na 2 substrate. Results are expressed as geometric mean of OD 405 values, subtracted with background, ± standard deviation of quadruplicate samples. Columns with same letter do not differ at 5% level of probability (Tukey HSD test).
    Figure Legend Snippet: A , Binding of decorin to recombinant Dbps in a Western blot assay. The upper panel represents SimplyBlue staining of the SDS-PAGE gel. The lower panel shows the adhesion of biotinylated decorin (1 μg/mL) to Dbps. B , Binding of decorin to recombinant Dbps in a microtiter plate assay. Recombinant Dbps (10 μg/mL) were attached on microtiter plates, and unspecific binding was blocked with BSA. The binding of biotinylated decorin (1 μg/mL) was detected with alkaline-phosphatase streptavidin and p-NPP-Na 2 substrate. Results are expressed as geometric mean of OD 405 values, subtracted with background, ± standard deviation of quadruplicate samples. Columns with same letter do not differ at 5% level of probability (Tukey HSD test).

    Techniques Used: Binding Assay, Recombinant, Western Blot, Staining, SDS Page, Standard Deviation

    Related Articles

    Enzyme-linked Immunosorbent Assay:

    Article Title: Elicitation of Neutralizing Antibodies Targeting the V2 Apex of the HIV Envelope Trimer in a Wild-Type Animal Model
    Article Snippet: .. ELISA SOSIP ELISA reagents were randomly biotinylated using a 2:1 molar ratio of biotin reagent to trimer using the EZ-link-NHS-PEG4 -Biotin kit (Thermo Fisher Scientific, 21324) MaxiSorp plates (Thermo Fisher Scientific) were coated overnight at 4°C with 2 μg/mL gp120 protein or 2 μg/mL streptavidin (Thermo Fisher Scientific). ..

    Article Title: Identification and characterization of a core fucosidase from the bacterium Elizabethkingia meningoseptica
    Article Snippet: .. IgE was detected by incubation with biotinylated anti-human IgE antibody (BioLegend, 325504) for 1 h at room temperature, followed by incubation with streptavidin-conjugated AP (Thermo Fisher Scientific, 21324) for 1 h. After one more round of washing, staining of ELISA plates was performed by incubation with 50 μl/well of AP substrate (Sigma, P7998). ..

    Staining:

    Article Title: Identification and characterization of a core fucosidase from the bacterium Elizabethkingia meningoseptica
    Article Snippet: .. IgE was detected by incubation with biotinylated anti-human IgE antibody (BioLegend, 325504) for 1 h at room temperature, followed by incubation with streptavidin-conjugated AP (Thermo Fisher Scientific, 21324) for 1 h. After one more round of washing, staining of ELISA plates was performed by incubation with 50 μl/well of AP substrate (Sigma, P7998). ..

    Incubation:

    Article Title: Identification and characterization of a core fucosidase from the bacterium Elizabethkingia meningoseptica
    Article Snippet: .. IgE was detected by incubation with biotinylated anti-human IgE antibody (BioLegend, 325504) for 1 h at room temperature, followed by incubation with streptavidin-conjugated AP (Thermo Fisher Scientific, 21324) for 1 h. After one more round of washing, staining of ELISA plates was performed by incubation with 50 μl/well of AP substrate (Sigma, P7998). ..

    Article Title: Dissociation of the Tubulin Dimer Is Extremely Slow, Thermodynamically Very Unfavorable, and Reversible in the Absence of an Energy Source
    Article Snippet: .. After three 10-minute washes in PBS, the membrane was incubated for 0.5–16 h with alkaline phosphatase–conjugated streptavidin (Cat no. 21324; Pierce Chemical , Rockford, IL) diluted 46,000-fold in PBS. .. After two 10-min washes with PBS and one with Tris-buffered saline the membrane was reacted with Amersham Pharmacia ECF reagent (Cat No. PRN5785), following the manufacturer's instructions.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher streptavidin conjugated to alkaline phosphatase
    Streptavidin Conjugated To Alkaline Phosphatase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin conjugated to alkaline phosphatase/product/Thermo Fisher
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    streptavidin conjugated to alkaline phosphatase - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Thermo Fisher streptavidin conjugated alkaline phosphatase sav alp
    Schematic illustration of tdEV sensing using a sandwich immunoassay and redox cycling on nIDEs resulting in a two-level selectivity and a two-level amplification. tdEVs are captured using C-AE tethered to electrodes (first level of selectivity). The binding of R-AE to the tdEVs completes the antibody–antigen-antibody sandwich (second-level selectivity), after which the enzyme <t>ALP</t> is introduced using a <t>biotin–SAV</t> interaction. ALP provides an enzymatic amplification of pAPP to pAP by substrate cleavage (first-level amplification), which is followed by an electrochemical signal amplification via the oxidation of pAP to pQI and subsequent redox cycling thereof between the nIDE electrodes (second-level amplification).
    Streptavidin Conjugated Alkaline Phosphatase Sav Alp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/streptavidin conjugated alkaline phosphatase sav alp/product/Thermo Fisher
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    streptavidin conjugated alkaline phosphatase sav alp - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    84
    prozyme alkaline phosphatase conjugated streptavidin sa ap
    Detection of PD-1 and PD-L1 expression on canine cell lines. (A) PD-1 expression on CLGL-90 cells was detected with JC053 hybridoma culture supernatant. Biotinylated anti-mouse IgA and FITC-conjugated <t>streptavidin</t> was used. Mouse IgA isotype control was used as negative control. Light gray is for isotype control and dark gray is for JC053. (B) Four purified anti-PD-L1s were compared in their capacity to bind activated DH82 cells. DH82 cells were cultivated either with or without 10ng/ml IFN-γ and JC071, JC173, JC194, and JC205 binding to PD-L1 was assessed. Histograms show unstimulated DH82 without anti-PD-L1 (light gray), unstimulated DH82 with anti-PD-L1 (medium gray), and IFN-γ stimulated DH82 with anti-PD-L1 (dark gray), respectively. FITC conjugated Anti-Mouse IgG was used with all samples including no anti-PD-L1 sample, as secondary antibody. PD-1 or PD-L1 positive population was selected following FSC, SSC gating, single cell gating, and live cell gating.
    Alkaline Phosphatase Conjugated Streptavidin Sa Ap, supplied by prozyme, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alkaline phosphatase conjugated streptavidin sa ap/product/prozyme
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alkaline phosphatase conjugated streptavidin sa ap - by Bioz Stars, 2020-09
    84/100 stars
      Buy from Supplier

    Image Search Results


    Schematic illustration of tdEV sensing using a sandwich immunoassay and redox cycling on nIDEs resulting in a two-level selectivity and a two-level amplification. tdEVs are captured using C-AE tethered to electrodes (first level of selectivity). The binding of R-AE to the tdEVs completes the antibody–antigen-antibody sandwich (second-level selectivity), after which the enzyme ALP is introduced using a biotin–SAV interaction. ALP provides an enzymatic amplification of pAPP to pAP by substrate cleavage (first-level amplification), which is followed by an electrochemical signal amplification via the oxidation of pAP to pQI and subsequent redox cycling thereof between the nIDE electrodes (second-level amplification).

    Journal: Nano Letters

    Article Title: Electrochemical Detection of Tumor-Derived Extracellular Vesicles on Nanointerdigitated Electrodes

    doi: 10.1021/acs.nanolett.9b02741

    Figure Lengend Snippet: Schematic illustration of tdEV sensing using a sandwich immunoassay and redox cycling on nIDEs resulting in a two-level selectivity and a two-level amplification. tdEVs are captured using C-AE tethered to electrodes (first level of selectivity). The binding of R-AE to the tdEVs completes the antibody–antigen-antibody sandwich (second-level selectivity), after which the enzyme ALP is introduced using a biotin–SAV interaction. ALP provides an enzymatic amplification of pAPP to pAP by substrate cleavage (first-level amplification), which is followed by an electrochemical signal amplification via the oxidation of pAP to pQI and subsequent redox cycling thereof between the nIDE electrodes (second-level amplification).

    Article Snippet: Streptavidin-conjugated alkaline phosphatase (SAV-ALP) was purchased from Thermo Fisher (Eindhoven, The Netherlands).

    Techniques: Amplification, Binding Assay

    Detection of PD-1 and PD-L1 expression on canine cell lines. (A) PD-1 expression on CLGL-90 cells was detected with JC053 hybridoma culture supernatant. Biotinylated anti-mouse IgA and FITC-conjugated streptavidin was used. Mouse IgA isotype control was used as negative control. Light gray is for isotype control and dark gray is for JC053. (B) Four purified anti-PD-L1s were compared in their capacity to bind activated DH82 cells. DH82 cells were cultivated either with or without 10ng/ml IFN-γ and JC071, JC173, JC194, and JC205 binding to PD-L1 was assessed. Histograms show unstimulated DH82 without anti-PD-L1 (light gray), unstimulated DH82 with anti-PD-L1 (medium gray), and IFN-γ stimulated DH82 with anti-PD-L1 (dark gray), respectively. FITC conjugated Anti-Mouse IgG was used with all samples including no anti-PD-L1 sample, as secondary antibody. PD-1 or PD-L1 positive population was selected following FSC, SSC gating, single cell gating, and live cell gating.

    Journal: PLoS ONE

    Article Title: Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function

    doi: 10.1371/journal.pone.0235518

    Figure Lengend Snippet: Detection of PD-1 and PD-L1 expression on canine cell lines. (A) PD-1 expression on CLGL-90 cells was detected with JC053 hybridoma culture supernatant. Biotinylated anti-mouse IgA and FITC-conjugated streptavidin was used. Mouse IgA isotype control was used as negative control. Light gray is for isotype control and dark gray is for JC053. (B) Four purified anti-PD-L1s were compared in their capacity to bind activated DH82 cells. DH82 cells were cultivated either with or without 10ng/ml IFN-γ and JC071, JC173, JC194, and JC205 binding to PD-L1 was assessed. Histograms show unstimulated DH82 without anti-PD-L1 (light gray), unstimulated DH82 with anti-PD-L1 (medium gray), and IFN-γ stimulated DH82 with anti-PD-L1 (dark gray), respectively. FITC conjugated Anti-Mouse IgG was used with all samples including no anti-PD-L1 sample, as secondary antibody. PD-1 or PD-L1 positive population was selected following FSC, SSC gating, single cell gating, and live cell gating.

    Article Snippet: The transferred membrane was blocked with 5% skim milk, incubated with alkaline phosphatase conjugated streptavidin (SA-AP), and developed using NBT/BCIP (Thermo Fisher Scientific) as a substrate.

    Techniques: Expressing, Negative Control, Purification, Binding Assay

    Evaluation of blocking activity by anti-canine PD-1 and PD-L1 antibodies. Antibodies were screened for their opposite ligand blocking capability. Hybridoma supernatants were added to ELISA plates coated with unbiotinylated PD-1Ig (A) or PD-L1Ig (B). After washing, biotinylated PD-L1Ig (A) or PD-1Ig (B) was used to detect blocking capacity of anti-PD-1 (A) and anti-PD-L1 (B) using HRP conjugated streptavidin to detect unblocked protein. All samples were assayed in duplicates. Each value is presented as mean ± standard deviation. In (A), JC053 value was significantly different from the rest of anti-PD-1 hybridomas with P value of 0.0002. In (B), JC071, JC173, JC194, and JC205 values were significantly different from PBS and medium values with P values less than 0.0001.

    Journal: PLoS ONE

    Article Title: Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function

    doi: 10.1371/journal.pone.0235518

    Figure Lengend Snippet: Evaluation of blocking activity by anti-canine PD-1 and PD-L1 antibodies. Antibodies were screened for their opposite ligand blocking capability. Hybridoma supernatants were added to ELISA plates coated with unbiotinylated PD-1Ig (A) or PD-L1Ig (B). After washing, biotinylated PD-L1Ig (A) or PD-1Ig (B) was used to detect blocking capacity of anti-PD-1 (A) and anti-PD-L1 (B) using HRP conjugated streptavidin to detect unblocked protein. All samples were assayed in duplicates. Each value is presented as mean ± standard deviation. In (A), JC053 value was significantly different from the rest of anti-PD-1 hybridomas with P value of 0.0002. In (B), JC071, JC173, JC194, and JC205 values were significantly different from PBS and medium values with P values less than 0.0001.

    Article Snippet: The transferred membrane was blocked with 5% skim milk, incubated with alkaline phosphatase conjugated streptavidin (SA-AP), and developed using NBT/BCIP (Thermo Fisher Scientific) as a substrate.

    Techniques: Blocking Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Expression of recombinant PD-1Ig and PD-L1Ig in D . melanogaster S2 cells. Expression of monomers was tested by Western blot. Biotinylated PD-1Ig and PD-L1Ig were expressed and secreted from D . melanogaster S2 cells. S2 cell culture supernatant expressing PD-1Ig (lane 2) or PD-L1Ig (lane 3) was analyzed against vector only control (lane 1) on Western blot in reducing condition. Alkaline phosphatase conjugated streptavidin was used to detect proteins.

    Journal: PLoS ONE

    Article Title: Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function

    doi: 10.1371/journal.pone.0235518

    Figure Lengend Snippet: Expression of recombinant PD-1Ig and PD-L1Ig in D . melanogaster S2 cells. Expression of monomers was tested by Western blot. Biotinylated PD-1Ig and PD-L1Ig were expressed and secreted from D . melanogaster S2 cells. S2 cell culture supernatant expressing PD-1Ig (lane 2) or PD-L1Ig (lane 3) was analyzed against vector only control (lane 1) on Western blot in reducing condition. Alkaline phosphatase conjugated streptavidin was used to detect proteins.

    Article Snippet: The transferred membrane was blocked with 5% skim milk, incubated with alkaline phosphatase conjugated streptavidin (SA-AP), and developed using NBT/BCIP (Thermo Fisher Scientific) as a substrate.

    Techniques: Expressing, Recombinant, Western Blot, Cell Culture, Plasmid Preparation

    Detection of PD-1 and PD-L1 expression on CHO cells expressing canine markers. Antibody specificity toward PD-1 and PD-L1 by anti-PD-1 (JC053) and anti-PD-L1s (JC071, JC173, JC194, and JC205) was tested on CHO cells by flow cytometry. (A) Anti-PD-1 and anti-PD-L1 bind to CHO-PD1 and CHO-PDL1, respectively, while they do not bind to untransfected CHO-K1. PD-1 positive population were selected following FSC, SSC gating, single cell gating, and live cell gating. PD-L1 positive population was selected following FSC, SSC gating, and single cell gating. (B) JC053, an anti-PD-1 antibody, was compared to a mouse IgA isotype control for staining CHO-PD1. Biotinylated anti-mouse IgA and APC conjugated streptavidin was used following primary antibody staining. A population of CHO-PD1 cells was selected on FSC and SSC gating. Single cells were selected and dead cells were excluded. Then, PD-1 positive cells are shown on histograms. Anti-PD-L1s were conjugated with PE and used to stain CHO-PDL1 cells. Appropriate isotype controls were included and anti-mouse IgG1 and anti-mouse IgG2a were used in combination with PE conjugated streptavidin. PE positive population is shown on histograms following FSC/SSC gating, single cell selection, and exclusion of dead cells.

    Journal: PLoS ONE

    Article Title: Development of canine PD-1/PD-L1 specific monoclonal antibodies and amplification of canine T cell function

    doi: 10.1371/journal.pone.0235518

    Figure Lengend Snippet: Detection of PD-1 and PD-L1 expression on CHO cells expressing canine markers. Antibody specificity toward PD-1 and PD-L1 by anti-PD-1 (JC053) and anti-PD-L1s (JC071, JC173, JC194, and JC205) was tested on CHO cells by flow cytometry. (A) Anti-PD-1 and anti-PD-L1 bind to CHO-PD1 and CHO-PDL1, respectively, while they do not bind to untransfected CHO-K1. PD-1 positive population were selected following FSC, SSC gating, single cell gating, and live cell gating. PD-L1 positive population was selected following FSC, SSC gating, and single cell gating. (B) JC053, an anti-PD-1 antibody, was compared to a mouse IgA isotype control for staining CHO-PD1. Biotinylated anti-mouse IgA and APC conjugated streptavidin was used following primary antibody staining. A population of CHO-PD1 cells was selected on FSC and SSC gating. Single cells were selected and dead cells were excluded. Then, PD-1 positive cells are shown on histograms. Anti-PD-L1s were conjugated with PE and used to stain CHO-PDL1 cells. Appropriate isotype controls were included and anti-mouse IgG1 and anti-mouse IgG2a were used in combination with PE conjugated streptavidin. PE positive population is shown on histograms following FSC/SSC gating, single cell selection, and exclusion of dead cells.

    Article Snippet: The transferred membrane was blocked with 5% skim milk, incubated with alkaline phosphatase conjugated streptavidin (SA-AP), and developed using NBT/BCIP (Thermo Fisher Scientific) as a substrate.

    Techniques: Expressing, Flow Cytometry, Staining, Selection