triethylammonium salt Thermo Fisher Search Results


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  • 90
    Thermo Fisher alexa fluor 488 azide
    Reaction scheme for creating click-conjugated dendrons employed in the in vitro studies. (i) glutaric anhydride, MeOH, 24 hrs, r.t. (ii) EDC, c(RGDyK), H 2 O/DMSO, 2 days, r.t. (iii) <t>Alexa</t> <t>Fluor</t> 488 azide OR biotin-dPEG 3+4 -azide OR MTX - azide (5) OP azide-G2(AF)
    Alexa Fluor 488 Azide, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 702 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Thermo Fisher tris triethylammonium salt
    Reaction scheme for creating click-conjugated dendrons employed in the in vitro studies. (i) glutaric anhydride, MeOH, 24 hrs, r.t. (ii) EDC, c(RGDyK), H 2 O/DMSO, 2 days, r.t. (iii) <t>Alexa</t> <t>Fluor</t> 488 azide OR biotin-dPEG 3+4 -azide OR MTX - azide (5) OP azide-G2(AF)
    Tris Triethylammonium Salt, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher triethylammonium salt tritc dhpe
    Analysis of domain-specific <t>TRITC-DHPE</t> diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p
    Triethylammonium Salt Tritc Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher triethylammonium salt rh pe
    Analysis of domain-specific <t>TRITC-DHPE</t> diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p
    Triethylammonium Salt Rh Pe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 76/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher nbd pe
    Time-lapse fluorescence microscopy images of shrinking vesicles. (a) The GFP leaked from DOPC/ 5 GUV upon the shrinkage of the vesicle by PLA 2 treatment. The GUV membrane was stained with Texas Red <t>DHPE.</t> (b) The 100 nm red fluorescent sulfate-modified polystyrene nanoparticles were contained and concentrated in DOPC/ 5 GUV as the vesicle shrank. The membrane was stained with <t>NBD-PE.</t>
    Nbd Pe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher alexa fluor 594
    Generation of antioxidant over-expressing parasites. Transgenic parasites were created in the RHΔhxgprt background strain using plasmids designed to over-express HA-tagged versions of glutaredoxin (Glut), catalase (Cat), or peroxiredoxin 2 (PRx2) under the control of the gra1 gene promoter. ( a ) Over-expression was verified using quantitative real-time PCR to measure transcript levels. ( b ) Western blot analysis was used to confirm protein production and that the resultant proteins were the appropriate size. ( c ) Immunofluorescence assays were performed for protein localization, utilizing a rabbit monoclonal antibody against the HA tag and an <t>Alexa</t> <t>Fluor</t> 594 secondary antibody. ( d ) General parasite replication was determined by doubling assay. Confluent HFF in 24-well plates were infected with the indicated parasite lines for 5 hours before the medium was replaced with fresh growth medium. Parasites were allowed to grow for 32 hours before being methanol-fixed and stained with DiffQuick. The number of individual parasites per vacuole was enumerated for at least 50 vacuoles. Data is presented as percent of vacuoles from 4 experiments. Student’s t-test was employed for statistical analysis. Scale bars equal 2 μm.
    Alexa Fluor 594, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 13743 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 594 alkyne alexa fluor 594 carboxamido 5 and 6 propargyl bis triethylammonium salt
    Images of immunofluorescence by confocal laser scanning microscopy (CLSM) of four different breast cancer cell lines: MCF-7, SKBR3, T47D and MDA-MB-231 ( a ) bright field; ( b ) staining of the cell nucleus by the fluorophore DAPI (blue); ( c ) indirect detection of HER2 by a primary anti-HER2 antibody and an anti-anti-HER2 secondary antibody bound to <t>Alexa</t> <t>Fluor</t> 594 fluorophore (red) and ( d ) cell localization of HER2 receptor by overlapping of images (DAPI + HER2). Magnification: 63×.
    Alexa Fluor 594 Alkyne Alexa Fluor 594 Carboxamido 5 And 6 Propargyl Bis Triethylammonium Salt, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 75/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher alexafluor 488
    (a) Flow analysis to show aggregated human γ-globulin (AHG)–AlexaFluor® 488 binding to CD4 +  T cells in purified peripheral blood mononuclear cells (PBMCs). Cells were gated using forward- and side-scatter. CD4 +  lymphocytes were
    Alexafluor 488, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 6999 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher oregon green 488 taxol
    Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 <t>Taxol;</t> WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.
    Oregon Green 488 Taxol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Thermo Fisher triethylammonium salt tr dppe
    Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 <t>Taxol;</t> WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.
    Triethylammonium Salt Tr Dppe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    75
    Thermo Fisher triethylammonium salt dansyl
    Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 <t>Taxol;</t> WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.
    Triethylammonium Salt Dansyl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher tris
    Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 <t>Taxol;</t> WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.
    Tris, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 11798 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher ped6
    The activated complex preferentially transfers sn-1 acyl chains from glycerophospholipids to cholesterol. ( A ) Enzymatic assays with liposomes containing two sn-1 and sn-2 selective fluorescent probes PEDA1 (●) and <t>PED6</t> (◆). SseJ does not cleave PEDA1 in absence of GTPgS loaded RhoA (❍). ( B ) TLC analysis of the fluorescent reaction products confirming the formation of a BODIPY ® modified cholesterol ester (CE) in presence of PEDA1. Bee venom phospholipase A2 (PLA2), which generates free fatty acids (FA) was used as a control.
    Ped6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 647 atp alexa fluor 647
    Properties of the fluorescent <t>kinesin-ATP</t> complex. ( a ) Emission spectra of <t>Alexa</t> Fluor 647-ATP: black squares show the emission spectrum of Alexa Fluor 647-ATP bound to Alexa Fluor 555-labeled kinesin (∼20 nM kinesin with an approximately equimolar
    Alexa Fluor 647 Atp Alexa Fluor 647, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher fluorescein dhpe
    Characterization of liposomal formulations. ( A ) Representative Cryo-TEM image of DLPC/Chol/Cholesteryl/PEG 600 -Chol (5∶3.5∶1∶0.5) liposomes extruded through a 200 nm pore size membrane. ( B ) Confocal fluorescence image of a single liposome tagged on its lipid bilayer with Marina <t>Blue-DHPE</t> (blue) and its corresponding fluorescence intensity profile. ( C ) Confocal fluorescence image of a single Marina Blue-labeled liposome containing AlexaFluor594-labeled <t>LPS</t> (red) and their corresponding fluorescence intensity profiles. ( D ) Confocal fluorescence image of a single Marina Blue-labeled liposome containing fluorescein-labeled poly (I:C) and their corresponding fluorescence intensity profiles. ( E ) Schematic representation of the liposomal IS-cocktail (NL c ) showing the presence of both encapsulated LPS (red) and poly (I:C) (green) in the lipidic bilayer of liposomes. ( F ) Confocal fluorescence image of a single liposome containing both fluorescein-labeled poly (I:C) (green) and AlexaFluor594-labeled LPS (red) and their corresponding fluorescence intensity profiles.
    Fluorescein Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher biotin x dhpe lipids
    Single <t>Biotin-X-DHPE</t> molecules tagged with Streptavidin-Alexa546 were tracked on the curved supported lipid bilayers (ROC = 28 nm) in space and time. ( a ) example trajectories (blue) show the heterogeneous dynamics observed on both flat (white) and curved (black) regions. Scale bar = 1 µm; ( b ) the average step a molecule takes over 0.228 s (5 frames) when starting at a region of curvature (white) or at a flat region (grey); ( c ) the distribution of steps observed at curved and flat regions (shown in Figure S3 ) was fitted to Equation 3 to obtain the diffusion coefficients and the percentage of steps moving at that rate. The average D is plotted for t = 0.091, 0.228 and 0.456. Note that there is no fast component for the tracks that start at regions of curvature.
    Biotin X Dhpe Lipids, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 1 2 dihexadecanoyl sn glycero 3 phosphoethanolamine triethylammonium salt texasred dhpe
    Single <t>Biotin-X-DHPE</t> molecules tagged with Streptavidin-Alexa546 were tracked on the curved supported lipid bilayers (ROC = 28 nm) in space and time. ( a ) example trajectories (blue) show the heterogeneous dynamics observed on both flat (white) and curved (black) regions. Scale bar = 1 µm; ( b ) the average step a molecule takes over 0.228 s (5 frames) when starting at a region of curvature (white) or at a flat region (grey); ( c ) the distribution of steps observed at curved and flat regions (shown in Figure S3 ) was fitted to Equation 3 to obtain the diffusion coefficients and the percentage of steps moving at that rate. The average D is plotted for t = 0.091, 0.228 and 0.456. Note that there is no fast component for the tracks that start at regions of curvature.
    1 2 Dihexadecanoyl Sn Glycero 3 Phosphoethanolamine Triethylammonium Salt Texasred Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher bodipy fl dhpe
    ( A ) Chemical structures of the neutral lipids 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE). ( B ) Fluorescence micrographs of CHO cells treated with liposomes containing DOPC ( left ) or DOPE ( right ) as neutral lipid, DOTAP as cationic lipid, and <t>BODIPY</t> <t>FL-DHPE</t> as fluorescent component (1/1/0.1 mol/mol). Scale bars, 20 µm.
    Bodipy Fl Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Thermo Fisher triethylammonium salt rhod dhpe
    ( A ) Chemical structures of the neutral lipids 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE). ( B ) Fluorescence micrographs of CHO cells treated with liposomes containing DOPC ( left ) or DOPE ( right ) as neutral lipid, DOTAP as cationic lipid, and <t>BODIPY</t> <t>FL-DHPE</t> as fluorescent component (1/1/0.1 mol/mol). Scale bars, 20 µm.
    Triethylammonium Salt Rhod Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 76/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher triethylammonium salt texasred dhpe
    ( A ) Chemical structures of the neutral lipids 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE). ( B ) Fluorescence micrographs of CHO cells treated with liposomes containing DOPC ( left ) or DOPE ( right ) as neutral lipid, DOTAP as cationic lipid, and <t>BODIPY</t> <t>FL-DHPE</t> as fluorescent component (1/1/0.1 mol/mol). Scale bars, 20 µm.
    Triethylammonium Salt Texasred Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher triethylammonium salt lr dhpe
    Schematic representation of a multimodal SPION-based biotinylated magnetoliposome. Abbreviations: SPIONs, superparamagnetic iron oxide nanoparticles; DSPG, 1,2-Distearoyl-sn-glycero-3-phospho-rac-glycerol, sodium salt; Biotin-PEG2000-DSPE, 1,2-distearoyl-sn-glycero-3-phosphoethanol-amine-N- [biotinyl (polyethylene glycol)2000]; <t>Rhodamine-DHPE,</t> Lissamine™ rhodamine B 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, <t>triethylammonium</t> salt.
    Triethylammonium Salt Lr Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 76/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher bodipy fl gdp
    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
    Bodipy Fl Gdp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 647 azide
    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
    Alexa Fluor 647 Azide, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 319 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa647
    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
    Alexa647, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1218 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor 647 alkyne
    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
    Alexa Fluor 647 Alkyne, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher solution • 1 mm fluorescent azide
    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
    Solution • 1 Mm Fluorescent Azide, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher amine reactive dye pacific orange
    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
    Amine Reactive Dye Pacific Orange, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
    Alexa Fluor 647 Hydrazide, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rhodamine b 1 2 dihexadecanoyl sn glycero 3 phosphoethanolamine triethylammonium salt rhodamine dhpe
    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
    Rhodamine B 1 2 Dihexadecanoyl Sn Glycero 3 Phosphoethanolamine Triethylammonium Salt Rhodamine Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher triethylammonium salt rhodamine dhpe
    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of <t>GDP,</t> GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM <t>BODIPY-FL-GDP</t> (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.
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    Image Search Results


    Reaction scheme for creating click-conjugated dendrons employed in the in vitro studies. (i) glutaric anhydride, MeOH, 24 hrs, r.t. (ii) EDC, c(RGDyK), H 2 O/DMSO, 2 days, r.t. (iii) Alexa Fluor 488 azide OR biotin-dPEG 3+4 -azide OR MTX - azide (5) OP azide-G2(AF)

    Journal: Bioconjugate chemistry

    Article Title: RGD Dendron bodies; synthetic avidity agents with defined and potentially interchangeable effector sites that can substitute for antibodies

    doi: 10.1021/bc900217h

    Figure Lengend Snippet: Reaction scheme for creating click-conjugated dendrons employed in the in vitro studies. (i) glutaric anhydride, MeOH, 24 hrs, r.t. (ii) EDC, c(RGDyK), H 2 O/DMSO, 2 days, r.t. (iii) Alexa Fluor 488 azide OR biotin-dPEG 3+4 -azide OR MTX - azide (5) OP azide-G2(AF)

    Article Snippet: Alexa Fluor 488 azide and Prolong Gold mounting agent were obtained from Invitrogen. c(RGDyK) peptide was obtained from Peptide International.

    Techniques: In Vitro

    Dendron conjugate AF-G3(COOH) 11.9 (RGD) 4.1 (4) where Alexa Fluor 488 is conjugated to the dendron focal point via a 1,3-dipolar cycloaddition. The targeting moiety, c(RGDyK) peptide, is conjugated to the carboxylated surface of the dendron.

    Journal: Bioconjugate chemistry

    Article Title: RGD Dendron bodies; synthetic avidity agents with defined and potentially interchangeable effector sites that can substitute for antibodies

    doi: 10.1021/bc900217h

    Figure Lengend Snippet: Dendron conjugate AF-G3(COOH) 11.9 (RGD) 4.1 (4) where Alexa Fluor 488 is conjugated to the dendron focal point via a 1,3-dipolar cycloaddition. The targeting moiety, c(RGDyK) peptide, is conjugated to the carboxylated surface of the dendron.

    Article Snippet: Alexa Fluor 488 azide and Prolong Gold mounting agent were obtained from Invitrogen. c(RGDyK) peptide was obtained from Peptide International.

    Techniques:

    Analysis of domain-specific TRITC-DHPE diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p

    Journal: Biophysical Journal

    Article Title: Changes in Cholesterol Level Alter Integrin Sequestration in Raft-Mimicking Lipid Mixtures

    doi: 10.1016/j.bpj.2017.11.005

    Figure Lengend Snippet: Analysis of domain-specific TRITC-DHPE diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p

    Article Snippet: The dye-labeled phospholipids N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadec-anoyl- sn -glycero-3-phosphoethanolamine, triethylammonium salt (NBD-DHPE) and N-(6-tetramethylrhodamine-thiocarbamoyl)-1, 2-dihexadecanayl- sn -glycero-3-phosphoethanolamine, triethylammonium salt (TRITC-DHPE) were obtained from Invitrogen (Carlsbad, CA).

    Techniques:

    Time-lapse fluorescence microscopy images of shrinking vesicles. (a) The GFP leaked from DOPC/ 5 GUV upon the shrinkage of the vesicle by PLA 2 treatment. The GUV membrane was stained with Texas Red DHPE. (b) The 100 nm red fluorescent sulfate-modified polystyrene nanoparticles were contained and concentrated in DOPC/ 5 GUV as the vesicle shrank. The membrane was stained with NBD-PE.

    Journal: Chemical Science

    Article Title: Rapid access to phospholipid analogs using thiol-yne chemistry access to phospholipid analogs using thiol-yne chemistry †Electronic supplementary information (ESI) available: Details of experimental procedures, lipid characterization data (NMR, HRMS, steady-state fluorescence anisotropy), HPLC analysis, fluorescence microscopy images, liposome shrinkage data in the form of AVI files. See DOI: 10.1039/c5sc00653hClick here for additional data file.

    doi: 10.1039/c5sc00653h

    Figure Lengend Snippet: Time-lapse fluorescence microscopy images of shrinking vesicles. (a) The GFP leaked from DOPC/ 5 GUV upon the shrinkage of the vesicle by PLA 2 treatment. The GUV membrane was stained with Texas Red DHPE. (b) The 100 nm red fluorescent sulfate-modified polystyrene nanoparticles were contained and concentrated in DOPC/ 5 GUV as the vesicle shrank. The membrane was stained with NBD-PE.

    Article Snippet: 0.5 μL of 100 mM Texas Red DHPE or NBD-PE (Invitrogen, Carlsbad, CA) was added to 100 μL vesicle suspension, which was then added to a 96-well plate.

    Techniques: Fluorescence, Microscopy, Proximity Ligation Assay, Staining, Modification

    Generation of antioxidant over-expressing parasites. Transgenic parasites were created in the RHΔhxgprt background strain using plasmids designed to over-express HA-tagged versions of glutaredoxin (Glut), catalase (Cat), or peroxiredoxin 2 (PRx2) under the control of the gra1 gene promoter. ( a ) Over-expression was verified using quantitative real-time PCR to measure transcript levels. ( b ) Western blot analysis was used to confirm protein production and that the resultant proteins were the appropriate size. ( c ) Immunofluorescence assays were performed for protein localization, utilizing a rabbit monoclonal antibody against the HA tag and an Alexa Fluor 594 secondary antibody. ( d ) General parasite replication was determined by doubling assay. Confluent HFF in 24-well plates were infected with the indicated parasite lines for 5 hours before the medium was replaced with fresh growth medium. Parasites were allowed to grow for 32 hours before being methanol-fixed and stained with DiffQuick. The number of individual parasites per vacuole was enumerated for at least 50 vacuoles. Data is presented as percent of vacuoles from 4 experiments. Student’s t-test was employed for statistical analysis. Scale bars equal 2 μm.

    Journal: Scientific Reports

    Article Title: Oxidative stress generated during monensin treatment contributes to altered Toxoplasma gondii mitochondrial function

    doi: 10.1038/srep22997

    Figure Lengend Snippet: Generation of antioxidant over-expressing parasites. Transgenic parasites were created in the RHΔhxgprt background strain using plasmids designed to over-express HA-tagged versions of glutaredoxin (Glut), catalase (Cat), or peroxiredoxin 2 (PRx2) under the control of the gra1 gene promoter. ( a ) Over-expression was verified using quantitative real-time PCR to measure transcript levels. ( b ) Western blot analysis was used to confirm protein production and that the resultant proteins were the appropriate size. ( c ) Immunofluorescence assays were performed for protein localization, utilizing a rabbit monoclonal antibody against the HA tag and an Alexa Fluor 594 secondary antibody. ( d ) General parasite replication was determined by doubling assay. Confluent HFF in 24-well plates were infected with the indicated parasite lines for 5 hours before the medium was replaced with fresh growth medium. Parasites were allowed to grow for 32 hours before being methanol-fixed and stained with DiffQuick. The number of individual parasites per vacuole was enumerated for at least 50 vacuoles. Data is presented as percent of vacuoles from 4 experiments. Student’s t-test was employed for statistical analysis. Scale bars equal 2 μm.

    Article Snippet: Primary antibodies were visualized using either Alexa Fluor 594 or Alexa Fluor 488 secondary antibodies (Life Technologies).

    Techniques: Expressing, Transgenic Assay, Over Expression, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Infection, Staining

    Images of immunofluorescence by confocal laser scanning microscopy (CLSM) of four different breast cancer cell lines: MCF-7, SKBR3, T47D and MDA-MB-231 ( a ) bright field; ( b ) staining of the cell nucleus by the fluorophore DAPI (blue); ( c ) indirect detection of HER2 by a primary anti-HER2 antibody and an anti-anti-HER2 secondary antibody bound to Alexa Fluor 594 fluorophore (red) and ( d ) cell localization of HER2 receptor by overlapping of images (DAPI + HER2). Magnification: 63×.

    Journal: Nanomaterials

    Article Title: Development of a Nanostructured Platform for Identifying HER2-Heterogeneity of Breast Cancer Cells by Surface-Enhanced Raman Scattering

    doi: 10.3390/nano8070549

    Figure Lengend Snippet: Images of immunofluorescence by confocal laser scanning microscopy (CLSM) of four different breast cancer cell lines: MCF-7, SKBR3, T47D and MDA-MB-231 ( a ) bright field; ( b ) staining of the cell nucleus by the fluorophore DAPI (blue); ( c ) indirect detection of HER2 by a primary anti-HER2 antibody and an anti-anti-HER2 secondary antibody bound to Alexa Fluor 594 fluorophore (red) and ( d ) cell localization of HER2 receptor by overlapping of images (DAPI + HER2). Magnification: 63×.

    Article Snippet: The following materials were used: Dulbecco’s modified eagle’s medium (DMEM), fetal bovine serum (FBS), RPMI medium, F12 medium, non-essential amino acids, l -glutamine, sodium pyruvate, nunclon 6-well multidish, Alexa Fluor 594 dye (secondary antibody) and Penicillin/Streptomycin (antibiotic/antifungal) were purchased from Thermo Fisher Scientific, Mexico City, Mexico.

    Techniques: Immunofluorescence, Confocal Laser Scanning Microscopy, Multiple Displacement Amplification, Staining

    (a) Flow analysis to show aggregated human γ-globulin (AHG)–AlexaFluor® 488 binding to CD4 +  T cells in purified peripheral blood mononuclear cells (PBMCs). Cells were gated using forward- and side-scatter. CD4 +  lymphocytes were

    Journal: Clinical and Experimental Immunology

    Article Title: Immune complexes and late complement proteins trigger activation of Syk tyrosine kinase in human CD4+ T cells

    doi: 10.1111/j.1365-2249.2011.04505.x

    Figure Lengend Snippet: (a) Flow analysis to show aggregated human γ-globulin (AHG)–AlexaFluor® 488 binding to CD4 + T cells in purified peripheral blood mononuclear cells (PBMCs). Cells were gated using forward- and side-scatter. CD4 + lymphocytes were

    Article Snippet: One mg of the AHG protein was labelled with AlexaFluor® 488 using the protein labelling kit, as per the manufacturer's protocol (Invitrogen).

    Techniques: Flow Cytometry, Binding Assay, Purification

    Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 Taxol; WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.

    Journal: Current Chemical Genomics

    Article Title: Development of a Cell-Based High-Throughput Assay to Screen for Inhibitors of Organic Anion Transporting Polypeptides 1B1 and 1B3

    doi: 10.2174/1875397301004010001

    Figure Lengend Snippet: Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 Taxol; WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.

    Article Snippet: Materials Cell culture reagents, fluorescein-methotrexate (FMTX), Alexa Fluor® 488 methotrexate (AMTX), and Oregon Green® 488 Taxol (Flutax-2) were purchased from Invitrogen (Carlsbad, CA).

    Techniques: Expressing

    The activated complex preferentially transfers sn-1 acyl chains from glycerophospholipids to cholesterol. ( A ) Enzymatic assays with liposomes containing two sn-1 and sn-2 selective fluorescent probes PEDA1 (●) and PED6 (◆). SseJ does not cleave PEDA1 in absence of GTPgS loaded RhoA (❍). ( B ) TLC analysis of the fluorescent reaction products confirming the formation of a BODIPY ® modified cholesterol ester (CE) in presence of PEDA1. Bee venom phospholipase A2 (PLA2), which generates free fatty acids (FA) was used as a control.

    Journal: Science signaling

    Article Title: Activation of a bacterial virulence protein by the GTPase RhoA

    doi: 10.1126/scisignal.2000430

    Figure Lengend Snippet: The activated complex preferentially transfers sn-1 acyl chains from glycerophospholipids to cholesterol. ( A ) Enzymatic assays with liposomes containing two sn-1 and sn-2 selective fluorescent probes PEDA1 (●) and PED6 (◆). SseJ does not cleave PEDA1 in absence of GTPgS loaded RhoA (❍). ( B ) TLC analysis of the fluorescent reaction products confirming the formation of a BODIPY ® modified cholesterol ester (CE) in presence of PEDA1. Bee venom phospholipase A2 (PLA2), which generates free fatty acids (FA) was used as a control.

    Article Snippet: Selectivity of SseJ for acyltransfer from the sn-1 or sn-2 position was determined by lipase assays with liposomes containing 120 μM cholesterol 60 μM DOPC and 10 μM oleic acid supplemented with either 5 μM PEDA1 or 5 μM PED6 (Invitrogen, CA).

    Techniques: Thin Layer Chromatography, Modification

    Properties of the fluorescent kinesin-ATP complex. ( a ) Emission spectra of Alexa Fluor 647-ATP: black squares show the emission spectrum of Alexa Fluor 647-ATP bound to Alexa Fluor 555-labeled kinesin (∼20 nM kinesin with an approximately equimolar

    Journal:

    Article Title: Alternating-Site Mechanism of Kinesin-1 Characterized by Single-Molecule FRET Using Fluorescent ATP Analogues

    doi: 10.1016/j.bpj.2009.02.073

    Figure Lengend Snippet: Properties of the fluorescent kinesin-ATP complex. ( a ) Emission spectra of Alexa Fluor 647-ATP: black squares show the emission spectrum of Alexa Fluor 647-ATP bound to Alexa Fluor 555-labeled kinesin (∼20 nM kinesin with an approximately equimolar

    Article Snippet: For fluorimetric FRET assays, the release was induced by 1 mM Alexa Fluor 647 ATP (Alexa Fluor 647 2′-(or-3′)-O-( N -2-aminoethyl)urethane), hexa(triethylammonium)) ( ; Invitrogen, Carlsbad, CA).

    Techniques: Labeling

    Characterization of liposomal formulations. ( A ) Representative Cryo-TEM image of DLPC/Chol/Cholesteryl/PEG 600 -Chol (5∶3.5∶1∶0.5) liposomes extruded through a 200 nm pore size membrane. ( B ) Confocal fluorescence image of a single liposome tagged on its lipid bilayer with Marina Blue-DHPE (blue) and its corresponding fluorescence intensity profile. ( C ) Confocal fluorescence image of a single Marina Blue-labeled liposome containing AlexaFluor594-labeled LPS (red) and their corresponding fluorescence intensity profiles. ( D ) Confocal fluorescence image of a single Marina Blue-labeled liposome containing fluorescein-labeled poly (I:C) and their corresponding fluorescence intensity profiles. ( E ) Schematic representation of the liposomal IS-cocktail (NL c ) showing the presence of both encapsulated LPS (red) and poly (I:C) (green) in the lipidic bilayer of liposomes. ( F ) Confocal fluorescence image of a single liposome containing both fluorescein-labeled poly (I:C) (green) and AlexaFluor594-labeled LPS (red) and their corresponding fluorescence intensity profiles.

    Journal: PLoS ONE

    Article Title: A Novel Liposome-Based Nanocarrier Loaded with an LPS-dsRNA Cocktail for Fish Innate Immune System Stimulation

    doi: 10.1371/journal.pone.0076338

    Figure Lengend Snippet: Characterization of liposomal formulations. ( A ) Representative Cryo-TEM image of DLPC/Chol/Cholesteryl/PEG 600 -Chol (5∶3.5∶1∶0.5) liposomes extruded through a 200 nm pore size membrane. ( B ) Confocal fluorescence image of a single liposome tagged on its lipid bilayer with Marina Blue-DHPE (blue) and its corresponding fluorescence intensity profile. ( C ) Confocal fluorescence image of a single Marina Blue-labeled liposome containing AlexaFluor594-labeled LPS (red) and their corresponding fluorescence intensity profiles. ( D ) Confocal fluorescence image of a single Marina Blue-labeled liposome containing fluorescein-labeled poly (I:C) and their corresponding fluorescence intensity profiles. ( E ) Schematic representation of the liposomal IS-cocktail (NL c ) showing the presence of both encapsulated LPS (red) and poly (I:C) (green) in the lipidic bilayer of liposomes. ( F ) Confocal fluorescence image of a single liposome containing both fluorescein-labeled poly (I:C) (green) and AlexaFluor594-labeled LPS (red) and their corresponding fluorescence intensity profiles.

    Article Snippet: MarinaBlue-DHPE, fluorescein-DHPE, LPS-AlexaFluor594, antibiotic/antimycotic solution, TrypLE Express, Cell Mask Deep Red, Hoechst 33342 and Superscript III reverse transcriptase were purchased from Invitrogen.

    Techniques: Transmission Electron Microscopy, Fluorescence, Labeling

    Single Biotin-X-DHPE molecules tagged with Streptavidin-Alexa546 were tracked on the curved supported lipid bilayers (ROC = 28 nm) in space and time. ( a ) example trajectories (blue) show the heterogeneous dynamics observed on both flat (white) and curved (black) regions. Scale bar = 1 µm; ( b ) the average step a molecule takes over 0.228 s (5 frames) when starting at a region of curvature (white) or at a flat region (grey); ( c ) the distribution of steps observed at curved and flat regions (shown in Figure S3 ) was fitted to Equation 3 to obtain the diffusion coefficients and the percentage of steps moving at that rate. The average D is plotted for t = 0.091, 0.228 and 0.456. Note that there is no fast component for the tracks that start at regions of curvature.

    Journal: Membranes

    Article Title: Single Lipid Molecule Dynamics on Supported Lipid Bilayers with Membrane Curvature

    doi: 10.3390/membranes7010015

    Figure Lengend Snippet: Single Biotin-X-DHPE molecules tagged with Streptavidin-Alexa546 were tracked on the curved supported lipid bilayers (ROC = 28 nm) in space and time. ( a ) example trajectories (blue) show the heterogeneous dynamics observed on both flat (white) and curved (black) regions. Scale bar = 1 µm; ( b ) the average step a molecule takes over 0.228 s (5 frames) when starting at a region of curvature (white) or at a flat region (grey); ( c ) the distribution of steps observed at curved and flat regions (shown in Figure S3 ) was fitted to Equation 3 to obtain the diffusion coefficients and the percentage of steps moving at that rate. The average D is plotted for t = 0.091, 0.228 and 0.456. Note that there is no fast component for the tracks that start at regions of curvature.

    Article Snippet: The dynamics of Biotin-X-DHPE lipids were detected by in situ labeling with Alexa Fluor 546 (Thermo Fisher) conjugated to streptavidin (Strep-546).

    Techniques: Diffusion-based Assay

    ( A ) Chemical structures of the neutral lipids 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE). ( B ) Fluorescence micrographs of CHO cells treated with liposomes containing DOPC ( left ) or DOPE ( right ) as neutral lipid, DOTAP as cationic lipid, and BODIPY FL-DHPE as fluorescent component (1/1/0.1 mol/mol). Scale bars, 20 µm.

    Journal: International Journal of Molecular Sciences

    Article Title: Deciphering the Functional Composition of Fusogenic Liposomes

    doi: 10.3390/ijms19020346

    Figure Lengend Snippet: ( A ) Chemical structures of the neutral lipids 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC) and 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE). ( B ) Fluorescence micrographs of CHO cells treated with liposomes containing DOPC ( left ) or DOPE ( right ) as neutral lipid, DOTAP as cationic lipid, and BODIPY FL-DHPE as fluorescent component (1/1/0.1 mol/mol). Scale bars, 20 µm.

    Article Snippet: The fluorescently labelled lipids N -(4,4-difluoro-5,7-dimethyl-4-bora-3 a ,4 a -diaza- s -indacene-3-propionyl)-1,2-dihexadecanoyl- sn -glycero-3-phosphoethanolamine (triethylammonium salt) (BODIPY FL-DHPE) and 2-(4,4-difluoro-5-methyl-4-bora-3 a ,4 a -diazas-indacene-3-dodecanoyl)-1-hexadecanoyl- sn -glycero-3-phosphocholine (βBODIPY-C12 HPC), and the lipid analogue DiI(C18)7 (DiR) were ordered from Thermo Scientifics (Eugene, OR, USA).

    Techniques: Fluorescence

    ( A ) Liposomal zeta potential (blue triangles) and fusion efficiency (black squares) at varying cationic lipid concentration. Bar indicate standard deviations. ( B ) Flow cytometry dot plots to determine the cellular uptake pathway and its efficiency. Liposomes always contained the aromatic tracer BODIPY FL-DHPE. Its monomer signal was detected in the green, its dimer signal in the red channels after incubation with Chinese hamster ovary cells (CHO). Endocytotic liposomal uptake resulted in a nearly equal dimer and monomer signals of the tracer, while a high monomer and a low dimer signal were detected in the case of membrane fusion.

    Journal: International Journal of Molecular Sciences

    Article Title: Deciphering the Functional Composition of Fusogenic Liposomes

    doi: 10.3390/ijms19020346

    Figure Lengend Snippet: ( A ) Liposomal zeta potential (blue triangles) and fusion efficiency (black squares) at varying cationic lipid concentration. Bar indicate standard deviations. ( B ) Flow cytometry dot plots to determine the cellular uptake pathway and its efficiency. Liposomes always contained the aromatic tracer BODIPY FL-DHPE. Its monomer signal was detected in the green, its dimer signal in the red channels after incubation with Chinese hamster ovary cells (CHO). Endocytotic liposomal uptake resulted in a nearly equal dimer and monomer signals of the tracer, while a high monomer and a low dimer signal were detected in the case of membrane fusion.

    Article Snippet: The fluorescently labelled lipids N -(4,4-difluoro-5,7-dimethyl-4-bora-3 a ,4 a -diaza- s -indacene-3-propionyl)-1,2-dihexadecanoyl- sn -glycero-3-phosphoethanolamine (triethylammonium salt) (BODIPY FL-DHPE) and 2-(4,4-difluoro-5-methyl-4-bora-3 a ,4 a -diazas-indacene-3-dodecanoyl)-1-hexadecanoyl- sn -glycero-3-phosphocholine (βBODIPY-C12 HPC), and the lipid analogue DiI(C18)7 (DiR) were ordered from Thermo Scientifics (Eugene, OR, USA).

    Techniques: Concentration Assay, Flow Cytometry, Cytometry, Incubation

    Importance of the aromatic component. ( A ) Fluorescence micrographs of CHO cells treated with liposomes containing BODIPY FL-DHPE as aromatic component in fusogenic liposomes (FL) at 0.01 (*) and 0.1 (**) mol/mol concentration as well as in endocytotic liposomes (EL) at the same concentrations (# and ##). Scale bars, 20 µm. ( B ) Dependence of fusion efficiency on dye concentration in FLs (black) and in ELs (red) determined by flow cytometry. The signal intensity median of the whole cell population was plotted vs. BODIPY FL-DHPE molar ratio to the cationic DOTAP amount in the liposomes (n Bodipy FL-DHPE /n DOTAP mol/mol). Measurement points with standard deviations are shown as squares (FL) and circles (EL), respectively. Lines represent linear fits. ( C ) Molecular structures of the chain labelled lipid βBODIPY-C 12 HPC, the head labelled lipid BODIPY FL-DPHE, and the lipophilic membrane dye DiR incorporated in FLs as fluorescent components. The aromatic molecular parts are coloured green and red, representing their spectral emissions. Results for βBODIPY-C 12 .

    Journal: International Journal of Molecular Sciences

    Article Title: Deciphering the Functional Composition of Fusogenic Liposomes

    doi: 10.3390/ijms19020346

    Figure Lengend Snippet: Importance of the aromatic component. ( A ) Fluorescence micrographs of CHO cells treated with liposomes containing BODIPY FL-DHPE as aromatic component in fusogenic liposomes (FL) at 0.01 (*) and 0.1 (**) mol/mol concentration as well as in endocytotic liposomes (EL) at the same concentrations (# and ##). Scale bars, 20 µm. ( B ) Dependence of fusion efficiency on dye concentration in FLs (black) and in ELs (red) determined by flow cytometry. The signal intensity median of the whole cell population was plotted vs. BODIPY FL-DHPE molar ratio to the cationic DOTAP amount in the liposomes (n Bodipy FL-DHPE /n DOTAP mol/mol). Measurement points with standard deviations are shown as squares (FL) and circles (EL), respectively. Lines represent linear fits. ( C ) Molecular structures of the chain labelled lipid βBODIPY-C 12 HPC, the head labelled lipid BODIPY FL-DPHE, and the lipophilic membrane dye DiR incorporated in FLs as fluorescent components. The aromatic molecular parts are coloured green and red, representing their spectral emissions. Results for βBODIPY-C 12 .

    Article Snippet: The fluorescently labelled lipids N -(4,4-difluoro-5,7-dimethyl-4-bora-3 a ,4 a -diaza- s -indacene-3-propionyl)-1,2-dihexadecanoyl- sn -glycero-3-phosphoethanolamine (triethylammonium salt) (BODIPY FL-DHPE) and 2-(4,4-difluoro-5-methyl-4-bora-3 a ,4 a -diazas-indacene-3-dodecanoyl)-1-hexadecanoyl- sn -glycero-3-phosphocholine (βBODIPY-C12 HPC), and the lipid analogue DiI(C18)7 (DiR) were ordered from Thermo Scientifics (Eugene, OR, USA).

    Techniques: Fluorescence, Concentration Assay, Flow Cytometry, Cytometry

    Liposomal fusion efficiency vs. neutral lipid component. Fusion efficiency of liposomes containing a neutral lipid component with different chain lengths and chain saturations as well as DOTAP and BODIPY FL-DHPE (1/1/0.1 mol/mol) was determined on CHO cells by flow cytometry. Symbols: saturated chains—filled symbols, unsaturated chains—open symbols, PCs—squares, PEs—triangles.

    Journal: International Journal of Molecular Sciences

    Article Title: Deciphering the Functional Composition of Fusogenic Liposomes

    doi: 10.3390/ijms19020346

    Figure Lengend Snippet: Liposomal fusion efficiency vs. neutral lipid component. Fusion efficiency of liposomes containing a neutral lipid component with different chain lengths and chain saturations as well as DOTAP and BODIPY FL-DHPE (1/1/0.1 mol/mol) was determined on CHO cells by flow cytometry. Symbols: saturated chains—filled symbols, unsaturated chains—open symbols, PCs—squares, PEs—triangles.

    Article Snippet: The fluorescently labelled lipids N -(4,4-difluoro-5,7-dimethyl-4-bora-3 a ,4 a -diaza- s -indacene-3-propionyl)-1,2-dihexadecanoyl- sn -glycero-3-phosphoethanolamine (triethylammonium salt) (BODIPY FL-DHPE) and 2-(4,4-difluoro-5-methyl-4-bora-3 a ,4 a -diazas-indacene-3-dodecanoyl)-1-hexadecanoyl- sn -glycero-3-phosphocholine (βBODIPY-C12 HPC), and the lipid analogue DiI(C18)7 (DiR) were ordered from Thermo Scientifics (Eugene, OR, USA).

    Techniques: Flow Cytometry, Cytometry

    ( A ) Chemical structures of the positively charged lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and the neutral lipid 1,2-dioleoyl-3-dimethylammonium-propane (DODAP). ( B ) Hydrodynamic size and ( C ) zeta potential distributions of liposomes containing DOPE/DOTAP/BODIPY FL-DHPE and DOPE/DODAP/BODIPY FL-DHPE (1/1/0.1 mol/mol). ( D ) Fluorescence and phase contrast micrographs of CHO cells after treatment with liposomes containing DOTAP or DODAP. Scale bars, 20 µm.

    Journal: International Journal of Molecular Sciences

    Article Title: Deciphering the Functional Composition of Fusogenic Liposomes

    doi: 10.3390/ijms19020346

    Figure Lengend Snippet: ( A ) Chemical structures of the positively charged lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and the neutral lipid 1,2-dioleoyl-3-dimethylammonium-propane (DODAP). ( B ) Hydrodynamic size and ( C ) zeta potential distributions of liposomes containing DOPE/DOTAP/BODIPY FL-DHPE and DOPE/DODAP/BODIPY FL-DHPE (1/1/0.1 mol/mol). ( D ) Fluorescence and phase contrast micrographs of CHO cells after treatment with liposomes containing DOTAP or DODAP. Scale bars, 20 µm.

    Article Snippet: The fluorescently labelled lipids N -(4,4-difluoro-5,7-dimethyl-4-bora-3 a ,4 a -diaza- s -indacene-3-propionyl)-1,2-dihexadecanoyl- sn -glycero-3-phosphoethanolamine (triethylammonium salt) (BODIPY FL-DHPE) and 2-(4,4-difluoro-5-methyl-4-bora-3 a ,4 a -diazas-indacene-3-dodecanoyl)-1-hexadecanoyl- sn -glycero-3-phosphocholine (βBODIPY-C12 HPC), and the lipid analogue DiI(C18)7 (DiR) were ordered from Thermo Scientifics (Eugene, OR, USA).

    Techniques: Fluorescence

    Schematic representation of a multimodal SPION-based biotinylated magnetoliposome. Abbreviations: SPIONs, superparamagnetic iron oxide nanoparticles; DSPG, 1,2-Distearoyl-sn-glycero-3-phospho-rac-glycerol, sodium salt; Biotin-PEG2000-DSPE, 1,2-distearoyl-sn-glycero-3-phosphoethanol-amine-N- [biotinyl (polyethylene glycol)2000]; Rhodamine-DHPE, Lissamine™ rhodamine B 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt.

    Journal: International Journal of Nanomedicine

    Article Title: Anti-?v?3 antibody guided three-step pretargeting approach using magnetoliposomes for molecular magnetic resonance imaging of breast cancer angiogenesis

    doi: 10.2147/IJN.S38678

    Figure Lengend Snippet: Schematic representation of a multimodal SPION-based biotinylated magnetoliposome. Abbreviations: SPIONs, superparamagnetic iron oxide nanoparticles; DSPG, 1,2-Distearoyl-sn-glycero-3-phospho-rac-glycerol, sodium salt; Biotin-PEG2000-DSPE, 1,2-distearoyl-sn-glycero-3-phosphoethanol-amine-N- [biotinyl (polyethylene glycol)2000]; Rhodamine-DHPE, Lissamine™ rhodamine B 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt.

    Article Snippet: Lissamine™ rhodamine B 1,2-dihexadecanoyl -sn-glycero-3- phosphoethanolamine, triethylammonium salt (Rho-DHPE) was from Life Technologies (Carlsbad, CA, USA).

    Techniques:

    E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of GDP, GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM BODIPY-FL-GDP (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.

    Journal: Nucleic Acids Research

    Article Title: eIF2β is critical for eIF5-mediated GDP-dissociation inhibitor activity and translational control

    doi: 10.1093/nar/gkw657

    Figure Lengend Snippet: E189K has a modest impact on eIF2–eIF2B interaction and activity. ( A ) Affinity ( K d ) of GDP, GTP and Met–tRNA i to purified WT and mutant (β E189K) apo–eIF2 complexes measured by monitoring the fluorescence intensity of 100 nM BODIPY-FL-GDP (left), 100 nM BODIPY-FL-GTP (middle) or 20 nM BOP-N-Met–tRNA i with 1 mM GTP (right). ( B ) Kinetics of BODIPY-FL-GDP release from preformed eIF2 complexes in the presence of different eIF2B concentrations. K 1/2 and K max values were determined from curve fitting y = [( K max × x)/( K 1/2 + x)] + c. ( C ) Western blotting of IP of Flag-eIF2β, Flag-E189K and an untagged control from cells showing its co-association with known binding proteins. Quantification of at least three repeats using Li-Cor fluorescent secondary antibodies ±SE. Student's t -test indicates significant reduction in eIF2B–eIF2 interactions ( P = 2.9 × 10 −6 ) with E189K (marked *). Other factors are not significantly altered. Tif5 is indicated with an arrow, ‘♦’ marks a non-specific band.

    Article Snippet: Steady state fluorescence To assay nucleotide affinity, fluorescence intensity of 100 nM of BODIPY-FL-GDP or BODIPY-FL-GTP (Thermo Fisher Scientific) in 180 μl of assay buffer (30 mM HEPES, 100 mM KCl, 10 mM MgCl2 , pH 7.4) was measured using a Fluoromax-4 spectrophotometer (Horiba) (490 nm excitation, 509 nm emission).

    Techniques: Activity Assay, Purification, Mutagenesis, Fluorescence, Western Blot, Binding Assay

    E189K antagonizes eIF5 GDI activity. ( A ) Kinetics of BODIPY-FL-GDP release from preformed purified WT and mutant (β E189K) eIF2•GDP complexes (20 nM) with varying concentrations of GST-eIF5 ± SD ( n = 3). Molar eIF2:GST–eIF5 protein ratios are shown. Asterisks (*) mark points with statistically significant difference to WT ( P

    Journal: Nucleic Acids Research

    Article Title: eIF2β is critical for eIF5-mediated GDP-dissociation inhibitor activity and translational control

    doi: 10.1093/nar/gkw657

    Figure Lengend Snippet: E189K antagonizes eIF5 GDI activity. ( A ) Kinetics of BODIPY-FL-GDP release from preformed purified WT and mutant (β E189K) eIF2•GDP complexes (20 nM) with varying concentrations of GST-eIF5 ± SD ( n = 3). Molar eIF2:GST–eIF5 protein ratios are shown. Asterisks (*) mark points with statistically significant difference to WT ( P

    Article Snippet: Steady state fluorescence To assay nucleotide affinity, fluorescence intensity of 100 nM of BODIPY-FL-GDP or BODIPY-FL-GTP (Thermo Fisher Scientific) in 180 μl of assay buffer (30 mM HEPES, 100 mM KCl, 10 mM MgCl2 , pH 7.4) was measured using a Fluoromax-4 spectrophotometer (Horiba) (490 nm excitation, 509 nm emission).

    Techniques: Activity Assay, Purification, Mutagenesis