protein concentrator  (Millipore)


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    Structured Review

    Millipore protein concentrator
    Protein Concentrator, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/protein concentrator/product/Millipore
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    protein concentrator - by Bioz Stars, 2020-04
    85/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Bacillus anthracis o-succinylbenzoyl-CoA synthetase: reaction kinetics and a novel inhibitor mimicking its reaction intermediate †
    Article Snippet: Paragraph title: Cloning, expression, and purification of OSB-CoA synthetase from B. anthracis ... Prior to storage at −80°C, the buffer was exchanged with 50 mM Tris-HCl (pH 7.5), and the protein was concentrated to about 100 µM using a protein concentrator (Millipore, Carrigtwohill, Co. Cork, Ireland).

    Purification:

    Article Title: Bacillus anthracis o-succinylbenzoyl-CoA synthetase: reaction kinetics and a novel inhibitor mimicking its reaction intermediate †
    Article Snippet: Paragraph title: Cloning, expression, and purification of OSB-CoA synthetase from B. anthracis ... Prior to storage at −80°C, the buffer was exchanged with 50 mM Tris-HCl (pH 7.5), and the protein was concentrated to about 100 µM using a protein concentrator (Millipore, Carrigtwohill, Co. Cork, Ireland).

    Concentration Assay:

    Article Title: Bacillus anthracis o-succinylbenzoyl-CoA synthetase: reaction kinetics and a novel inhibitor mimicking its reaction intermediate †
    Article Snippet: Fractions containing OSB-CoA synthetase were pooled, and 3.8 M ammonium sulfate was added to a final concentration of 1 M. The resulting solution was loaded on a HP Phenyl Sepharose column (1.6 × 20) (Amersham Biosciences, Piscataway, NJ) equilibrated with Buffer C (50 mM Tris-HCl, 1 M ammonium sulfate, 0.2 M NaCl, pH 7.5). .. Prior to storage at −80°C, the buffer was exchanged with 50 mM Tris-HCl (pH 7.5), and the protein was concentrated to about 100 µM using a protein concentrator (Millipore, Carrigtwohill, Co. Cork, Ireland).

    Expressing:

    Article Title: Bacillus anthracis o-succinylbenzoyl-CoA synthetase: reaction kinetics and a novel inhibitor mimicking its reaction intermediate †
    Article Snippet: Paragraph title: Cloning, expression, and purification of OSB-CoA synthetase from B. anthracis ... Prior to storage at −80°C, the buffer was exchanged with 50 mM Tris-HCl (pH 7.5), and the protein was concentrated to about 100 µM using a protein concentrator (Millipore, Carrigtwohill, Co. Cork, Ireland).

    SDS Page:

    Article Title: Bacillus anthracis o-succinylbenzoyl-CoA synthetase: reaction kinetics and a novel inhibitor mimicking its reaction intermediate †
    Article Snippet: Fractions were analyzed by SDS-PAGE and those containing pure OSB-CoA synthetase were pooled. .. Prior to storage at −80°C, the buffer was exchanged with 50 mM Tris-HCl (pH 7.5), and the protein was concentrated to about 100 µM using a protein concentrator (Millipore, Carrigtwohill, Co. Cork, Ireland).

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  • 95
    Millipore β actin protein
    mRNA expression in callus tissue of control and PDLLA treated rats 2, 4 and 6 weeks post-surgery. Expression level of (A) Runx2, (B) HIF2A, (C) Wnt4, (D) Wnt10b, (E) GSK-3β, (F) β-catenin and (G) TCF-1. <t>Β-actin</t> is a control. Values are presented as mean ± standard deviation, *P
    β Actin Protein, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/β actin protein/product/Millipore
    Average 95 stars, based on 48 article reviews
    Price from $9.99 to $1999.99
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    92
    Millipore concentrated tr3 protein stocks
    Spatial determinants define the phenotypic properties of mesothelin-targeted <t>TR3.</t> ( a ) Schematic representation of the Meso/DAF fusion protein. The GPI-anchored human decay-accelerating factor (DAF) was used to physically elevate the mesothelin binding site of scFv-SS away from the plasma membrane. The short consensus repeats (SCRs) of DAF are indicated (1–4). ( b ) Western blot analysis of Jurkat cells stably expressing the mesothelin/DAF fusion protein (Meso/DAF, ~110 kDa). ( c ) Jurkat-Meso/DAF cells (~5%) were treated with increasing concentrations of TR3 and SS-TR3 and cell viability was determined. (n = 3, mean ± SEM; * P
    Concentrated Tr3 Protein Stocks, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/concentrated tr3 protein stocks/product/Millipore
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    concentrated tr3 protein stocks - by Bioz Stars, 2020-04
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    94
    Millipore immunoblot analysis protein concentration
    SDS-PAGE ( A ) and <t>immunoblot</t> ( B ) analyses of recombinant Tb4CL4-like. (A): Lane 1, uninduced protein; lane 2, induced protein; lane 3, flow-through of protein sample; lane 4, elution of equilibration buffer; lanes 5–9, elution buffer from 20, 40, 100, 120 and 140 mM imidazole buffer, respectively. (B): Lane 1, Tb4CL4-like-His fusion protein was analyzed by immunoblotting with anti-His as the primary antibody. The target band is marked in red. The positions and sizes (in kDa) of the weight standards in lane M are indicated.
    Immunoblot Analysis Protein Concentration, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immunoblot analysis protein concentration/product/Millipore
    Average 94 stars, based on 1 article reviews
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    immunoblot analysis protein concentration - by Bioz Stars, 2020-04
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    Image Search Results


    mRNA expression in callus tissue of control and PDLLA treated rats 2, 4 and 6 weeks post-surgery. Expression level of (A) Runx2, (B) HIF2A, (C) Wnt4, (D) Wnt10b, (E) GSK-3β, (F) β-catenin and (G) TCF-1. Β-actin is a control. Values are presented as mean ± standard deviation, *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Increased Runx2 expression associated with enhanced Wnt signaling in PDLLA internal fixation for fracture treatment

    doi: 10.3892/etm.2017.4216

    Figure Lengend Snippet: mRNA expression in callus tissue of control and PDLLA treated rats 2, 4 and 6 weeks post-surgery. Expression level of (A) Runx2, (B) HIF2A, (C) Wnt4, (D) Wnt10b, (E) GSK-3β, (F) β-catenin and (G) TCF-1. Β-actin is a control. Values are presented as mean ± standard deviation, *P

    Article Snippet: The β-actin protein (primary antibody from EMD Millipore, working concentration: 1:1,000) was blotted in the same membrane as an internal control to normalize the relative density.

    Techniques: Expressing, Standard Deviation

    Protein expression in callus tissue of control and PDLLA treated rats 2, 4 and 6 weeks post-surgery. Expression level of (A) Runx2, (B) HIF2A, (C) Wnt4, (D) Wnt10b, (E) GSK-3β, (F) β-catenin and (G) TCF-1 under PDLLA intramedullary fixation (PDLLA group) compared to that of Kirschner wire (control group). Β-actin is a control. Values are presented as mean ± standard deviation, *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Increased Runx2 expression associated with enhanced Wnt signaling in PDLLA internal fixation for fracture treatment

    doi: 10.3892/etm.2017.4216

    Figure Lengend Snippet: Protein expression in callus tissue of control and PDLLA treated rats 2, 4 and 6 weeks post-surgery. Expression level of (A) Runx2, (B) HIF2A, (C) Wnt4, (D) Wnt10b, (E) GSK-3β, (F) β-catenin and (G) TCF-1 under PDLLA intramedullary fixation (PDLLA group) compared to that of Kirschner wire (control group). Β-actin is a control. Values are presented as mean ± standard deviation, *P

    Article Snippet: The β-actin protein (primary antibody from EMD Millipore, working concentration: 1:1,000) was blotted in the same membrane as an internal control to normalize the relative density.

    Techniques: Expressing, Standard Deviation

    Spatial determinants define the phenotypic properties of mesothelin-targeted TR3. ( a ) Schematic representation of the Meso/DAF fusion protein. The GPI-anchored human decay-accelerating factor (DAF) was used to physically elevate the mesothelin binding site of scFv-SS away from the plasma membrane. The short consensus repeats (SCRs) of DAF are indicated (1–4). ( b ) Western blot analysis of Jurkat cells stably expressing the mesothelin/DAF fusion protein (Meso/DAF, ~110 kDa). ( c ) Jurkat-Meso/DAF cells (~5%) were treated with increasing concentrations of TR3 and SS-TR3 and cell viability was determined. (n = 3, mean ± SEM; * P

    Journal: Scientific Reports

    Article Title: Membrane-proximal TRAIL species are incapable of inducing short circuit apoptosis signaling: Implications for drug development and basic cytokine biology

    doi: 10.1038/srep22661

    Figure Lengend Snippet: Spatial determinants define the phenotypic properties of mesothelin-targeted TR3. ( a ) Schematic representation of the Meso/DAF fusion protein. The GPI-anchored human decay-accelerating factor (DAF) was used to physically elevate the mesothelin binding site of scFv-SS away from the plasma membrane. The short consensus repeats (SCRs) of DAF are indicated (1–4). ( b ) Western blot analysis of Jurkat cells stably expressing the mesothelin/DAF fusion protein (Meso/DAF, ~110 kDa). ( c ) Jurkat-Meso/DAF cells (~5%) were treated with increasing concentrations of TR3 and SS-TR3 and cell viability was determined. (n = 3, mean ± SEM; * P

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, MD).

    Techniques: Binding Assay, Western Blot, Stable Transfection, Expressing

    Mesothelin-targeted TR3 protects its target cells from cell death. ( a ) Representative example of Jurkat-Meso cell enrichment following SS-TR3 drug exposure. The cells were treated with SS-TR3 for six days or with medium alone (ctrl), followed by assessing the percentage of mesothelin-positive cells by flow cytometry. ( b ) Graphic representation of the data shown in ( a ) (n = 3, mean ± SEM; **** P

    Journal: Scientific Reports

    Article Title: Membrane-proximal TRAIL species are incapable of inducing short circuit apoptosis signaling: Implications for drug development and basic cytokine biology

    doi: 10.1038/srep22661

    Figure Lengend Snippet: Mesothelin-targeted TR3 protects its target cells from cell death. ( a ) Representative example of Jurkat-Meso cell enrichment following SS-TR3 drug exposure. The cells were treated with SS-TR3 for six days or with medium alone (ctrl), followed by assessing the percentage of mesothelin-positive cells by flow cytometry. ( b ) Graphic representation of the data shown in ( a ) (n = 3, mean ± SEM; **** P

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, MD).

    Techniques: Flow Cytometry, Cytometry

    Membrane-proximal TRAIL is unable to interact with its receptors on the same plasma cell membrane. ( a ) Schematic representation of the structural features of wild-type TRAIL, and the GPI-anchored TR3 forms TR3-GPI and TR3-DAF expressed from a bicistronic configuration using an eYFP marker. The short consensus repeats (SCRs) of DAF are indicated (1–4). ( b ) HEK293T cells express death receptor 5 (DR5). ( c ) HEK293T cells were transiently transfected with wt TRAIL, TR3-GPI and TR3-DAF and analyzed by flow cytometry using a blocking anti-TRAIL mAb. The expression profiles for wt TRAIL and TR3-GPI result in a diagonal pattern (proportional relationship between TRAIL and YFP marker), whereas TR3-DAF expression results in a lack of TRAIL detection in an area where DR5 expression was detected. ( d ) Chinese hamster ovary cells (DR5-), expressing human CAR receptor (CHO-CAR), were infected with adenoviral vectors containing TR3-GPI and TR3-DAF using the same bicistronic configuration described in ( a ) and analyzed for their TRAIL expression as described in ( c ). Both surface proteins produced similar expression profiles (proportional relationship between TRAIL and YFP marker) with TR3-DAF being 100% detectable over the entire YFP expression range, suggesting a lack of interference with a surface receptor for the TR3 domain of the fusion protein.

    Journal: Scientific Reports

    Article Title: Membrane-proximal TRAIL species are incapable of inducing short circuit apoptosis signaling: Implications for drug development and basic cytokine biology

    doi: 10.1038/srep22661

    Figure Lengend Snippet: Membrane-proximal TRAIL is unable to interact with its receptors on the same plasma cell membrane. ( a ) Schematic representation of the structural features of wild-type TRAIL, and the GPI-anchored TR3 forms TR3-GPI and TR3-DAF expressed from a bicistronic configuration using an eYFP marker. The short consensus repeats (SCRs) of DAF are indicated (1–4). ( b ) HEK293T cells express death receptor 5 (DR5). ( c ) HEK293T cells were transiently transfected with wt TRAIL, TR3-GPI and TR3-DAF and analyzed by flow cytometry using a blocking anti-TRAIL mAb. The expression profiles for wt TRAIL and TR3-GPI result in a diagonal pattern (proportional relationship between TRAIL and YFP marker), whereas TR3-DAF expression results in a lack of TRAIL detection in an area where DR5 expression was detected. ( d ) Chinese hamster ovary cells (DR5-), expressing human CAR receptor (CHO-CAR), were infected with adenoviral vectors containing TR3-GPI and TR3-DAF using the same bicistronic configuration described in ( a ) and analyzed for their TRAIL expression as described in ( c ). Both surface proteins produced similar expression profiles (proportional relationship between TRAIL and YFP marker) with TR3-DAF being 100% detectable over the entire YFP expression range, suggesting a lack of interference with a surface receptor for the TR3 domain of the fusion protein.

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, MD).

    Techniques: Marker, Transfection, Flow Cytometry, Cytometry, Blocking Assay, Expressing, Infection, Produced

    Membrane-anchored recombinant TR3 variants convert HEK293T cells into potent apoptosis-inducing effector cells. ( a ) Structural features of wild-type, native TRAIL (wt TRAIL) and recombinant TR3 forms anchored to the membrane via carboxyl-terminal incorporation of a VEGFR-2-derived transmembrane domain (TR3-TM), the GPI-encoding signal sequence of human DAF (TR3-GPI) or the entire mature form of human DAF (TR3-DAF). The short consensus repeats (SCRs) of DAF are indicated (1–4). Please note the distinct architectural differences between native TRAIL and the TR3 variants. In order for native TRAIL to be functionally active, three individual monomers have to join at the plasma membrane (1:1 stoichiometry), while each TR3 trimer associates with the membrane in a 3:1 stoichiometry (1 functional TR3 trimer/1 membrane anchor [TM or GPI]). ( b ) Schematic illustration of a bicistronic expression format 32 , in which the expression level of a membrane-anchored surface molecule is proportionally correlated with the expression pattern of the marker protein. ( c ) HEK293T cells were transiently transfected with TR3-DAF (green). The following day, the cells were washed and mixed with PKH26-labeled Jurkat target cells (red, E/T ratio 1:1.3) and allowed to sediment for 2 h at ambient temperature. The cells were then subjected to fluorescent microscopy. Please note the formation of heterotypic cell aggregates and smooth cell-cell junctions, indicative of tight interactions between TR3-DAF-expressing 293T cells and DR5-positive Jurkat cells. ( d ) Transiently transfected HEK293T cells were overlaid with Jurkat cells for 24 h, the mixtures were harvested and subjected to FACS analysis (FSC/SSC gating). Non-transfected HEK293T cells were used as a control. Please note that the viable Jurkat cells completely disappeared from their respective analysis gates following exposure to TR3-GPI- and TR3-DAF-expressing HEK293T cells. The “Debris” gate contains HEK293T cellular debris at baseline (upper panels) and additional debris originating from the “Jurkat” gates only under conditions in which HEK293T cells express the membrane-anchored TR3 variants TR3GPI and TR3DAF (lower panels). ( e ) Quantitative analysis of the data presented in ( d ) after background subtraction (n = 3, **** P

    Journal: Scientific Reports

    Article Title: Membrane-proximal TRAIL species are incapable of inducing short circuit apoptosis signaling: Implications for drug development and basic cytokine biology

    doi: 10.1038/srep22661

    Figure Lengend Snippet: Membrane-anchored recombinant TR3 variants convert HEK293T cells into potent apoptosis-inducing effector cells. ( a ) Structural features of wild-type, native TRAIL (wt TRAIL) and recombinant TR3 forms anchored to the membrane via carboxyl-terminal incorporation of a VEGFR-2-derived transmembrane domain (TR3-TM), the GPI-encoding signal sequence of human DAF (TR3-GPI) or the entire mature form of human DAF (TR3-DAF). The short consensus repeats (SCRs) of DAF are indicated (1–4). Please note the distinct architectural differences between native TRAIL and the TR3 variants. In order for native TRAIL to be functionally active, three individual monomers have to join at the plasma membrane (1:1 stoichiometry), while each TR3 trimer associates with the membrane in a 3:1 stoichiometry (1 functional TR3 trimer/1 membrane anchor [TM or GPI]). ( b ) Schematic illustration of a bicistronic expression format 32 , in which the expression level of a membrane-anchored surface molecule is proportionally correlated with the expression pattern of the marker protein. ( c ) HEK293T cells were transiently transfected with TR3-DAF (green). The following day, the cells were washed and mixed with PKH26-labeled Jurkat target cells (red, E/T ratio 1:1.3) and allowed to sediment for 2 h at ambient temperature. The cells were then subjected to fluorescent microscopy. Please note the formation of heterotypic cell aggregates and smooth cell-cell junctions, indicative of tight interactions between TR3-DAF-expressing 293T cells and DR5-positive Jurkat cells. ( d ) Transiently transfected HEK293T cells were overlaid with Jurkat cells for 24 h, the mixtures were harvested and subjected to FACS analysis (FSC/SSC gating). Non-transfected HEK293T cells were used as a control. Please note that the viable Jurkat cells completely disappeared from their respective analysis gates following exposure to TR3-GPI- and TR3-DAF-expressing HEK293T cells. The “Debris” gate contains HEK293T cellular debris at baseline (upper panels) and additional debris originating from the “Jurkat” gates only under conditions in which HEK293T cells express the membrane-anchored TR3 variants TR3GPI and TR3DAF (lower panels). ( e ) Quantitative analysis of the data presented in ( d ) after background subtraction (n = 3, **** P

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, MD).

    Techniques: Recombinant, Derivative Assay, Sequencing, Functional Assay, Expressing, Marker, Transfection, Labeling, Microscopy, FACS

    Reciprocal polarities are required for a functional interaction between ligand and receptor of the TNF superfamily member TRAIL. ( a ) Experimental evidence suggests that the mesothelin-targeted SS-TR3 is incapable of inducing apoptosis on cells it binds, represented by a state of “equal polarity” between ligand and receptor. Only an elongated spacer, placed either into the cancer drug itself (SS-S-TR3) or being part of the target antigen (Meso/DAF) that lifts the mesothelin binding site of SS-TR3 away from the membrane, allows formation of a “reciprocal polarity”, a requirement for a functional interaction between ligand and receptors. ( b ) Both mesothelin-targeted TR3 variants are capable of inducing death of bystander cells. ( c ) The phenotypic similarity of native TRAIL with its GPI-anchored variant TR3-GPI prevents apoptosis induction on cells that express both the ligand and the receptor. The “spacer-containing” fusion protein TR3-DAF differs from the latter variants due to the elevated location of the TR3 trimer relative to the plasma membrane. The short consensus repeats (SCRs) of the spacer (SCR1 of DAF and SCRs 15–17 of CR1) as well as DAF (1–4) are indicated for clarity.

    Journal: Scientific Reports

    Article Title: Membrane-proximal TRAIL species are incapable of inducing short circuit apoptosis signaling: Implications for drug development and basic cytokine biology

    doi: 10.1038/srep22661

    Figure Lengend Snippet: Reciprocal polarities are required for a functional interaction between ligand and receptor of the TNF superfamily member TRAIL. ( a ) Experimental evidence suggests that the mesothelin-targeted SS-TR3 is incapable of inducing apoptosis on cells it binds, represented by a state of “equal polarity” between ligand and receptor. Only an elongated spacer, placed either into the cancer drug itself (SS-S-TR3) or being part of the target antigen (Meso/DAF) that lifts the mesothelin binding site of SS-TR3 away from the membrane, allows formation of a “reciprocal polarity”, a requirement for a functional interaction between ligand and receptors. ( b ) Both mesothelin-targeted TR3 variants are capable of inducing death of bystander cells. ( c ) The phenotypic similarity of native TRAIL with its GPI-anchored variant TR3-GPI prevents apoptosis induction on cells that express both the ligand and the receptor. The “spacer-containing” fusion protein TR3-DAF differs from the latter variants due to the elevated location of the TR3 trimer relative to the plasma membrane. The short consensus repeats (SCRs) of the spacer (SCR1 of DAF and SCRs 15–17 of CR1) as well as DAF (1–4) are indicated for clarity.

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, MD).

    Techniques: Functional Assay, Binding Assay, Variant Assay

    Mesothelin-targeting increases the bioactivity of soluble TR3. ( a ) Schematic representation of recombinant TR3 forms. The functional domain of secreted wild-type TRAIL has been genetically linked, resulting in the TRAIL-based drug platform TR3 (left). TR3 was subsequently fused at its N-terminus with scFv-SS, resulting in the mesothelin-targeted TR3 variant SS-TR3 (right). ( b ) Western blot analysis of recombinant TR3 forms produced in HEK293T cells (anti-TRAIL pAb). Supernatant from mock transfected cells was used as a control (lane 1). ( c ) Schematic representation of FLAG-tagged, mature human mesothelin inserted into the membrane via a glycosylphosphatidyl (GPI) anchor. Western blot analysis of Jurkat cells infected with a retroviral vector encoding FLAG-tagged human mesothelin (~40 kDa). ( d ) Anti-FLAG immunostaining confirms expression of the biomarker on Jurkat cells using flow cytometry. ( e ) Jurkat-WT and ( f ) ~5% Jurkat-Meso cells were treated with increasing concentrations of TR3 and SS-TR3 and cell viability was determined (n = 3, mean ± SEM; ns, not significant, * P

    Journal: Scientific Reports

    Article Title: Membrane-proximal TRAIL species are incapable of inducing short circuit apoptosis signaling: Implications for drug development and basic cytokine biology

    doi: 10.1038/srep22661

    Figure Lengend Snippet: Mesothelin-targeting increases the bioactivity of soluble TR3. ( a ) Schematic representation of recombinant TR3 forms. The functional domain of secreted wild-type TRAIL has been genetically linked, resulting in the TRAIL-based drug platform TR3 (left). TR3 was subsequently fused at its N-terminus with scFv-SS, resulting in the mesothelin-targeted TR3 variant SS-TR3 (right). ( b ) Western blot analysis of recombinant TR3 forms produced in HEK293T cells (anti-TRAIL pAb). Supernatant from mock transfected cells was used as a control (lane 1). ( c ) Schematic representation of FLAG-tagged, mature human mesothelin inserted into the membrane via a glycosylphosphatidyl (GPI) anchor. Western blot analysis of Jurkat cells infected with a retroviral vector encoding FLAG-tagged human mesothelin (~40 kDa). ( d ) Anti-FLAG immunostaining confirms expression of the biomarker on Jurkat cells using flow cytometry. ( e ) Jurkat-WT and ( f ) ~5% Jurkat-Meso cells were treated with increasing concentrations of TR3 and SS-TR3 and cell viability was determined (n = 3, mean ± SEM; ns, not significant, * P

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, MD).

    Techniques: Recombinant, Functional Assay, Variant Assay, Western Blot, Produced, Transfection, Infection, Plasmid Preparation, Immunostaining, Expressing, Biomarker Assay, Flow Cytometry, Cytometry

    TR3 has improved pharmacological characteristics in vitro and in vivo

    Journal: Molecular cancer therapeutics

    Article Title: A genetically-encoded multifunctional TRAIL trimer facilitates cell-specific targeting and tumor cell killing

    doi: 10.1158/1535-7163.MCT-10-0225

    Figure Lengend Snippet: TR3 has improved pharmacological characteristics in vitro and in vivo

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, Maryland).

    Techniques: In Vitro, In Vivo

    TR3 is a powerful inducer of apoptosis

    Journal: Molecular cancer therapeutics

    Article Title: A genetically-encoded multifunctional TRAIL trimer facilitates cell-specific targeting and tumor cell killing

    doi: 10.1158/1535-7163.MCT-10-0225

    Figure Lengend Snippet: TR3 is a powerful inducer of apoptosis

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, Maryland).

    Techniques:

    RBC-targeted TR3 kills tumor cells in vitro

    Journal: Molecular cancer therapeutics

    Article Title: A genetically-encoded multifunctional TRAIL trimer facilitates cell-specific targeting and tumor cell killing

    doi: 10.1158/1535-7163.MCT-10-0225

    Figure Lengend Snippet: RBC-targeted TR3 kills tumor cells in vitro

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, Maryland).

    Techniques: In Vitro

    TR3-coated RBCs attach to death receptor-positive target cells

    Journal: Molecular cancer therapeutics

    Article Title: A genetically-encoded multifunctional TRAIL trimer facilitates cell-specific targeting and tumor cell killing

    doi: 10.1158/1535-7163.MCT-10-0225

    Figure Lengend Snippet: TR3-coated RBCs attach to death receptor-positive target cells

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, Maryland).

    Techniques:

    Human pancreatic cancer cells are killed in vivo following coinjection with TR3-coated RBCs

    Journal: Molecular cancer therapeutics

    Article Title: A genetically-encoded multifunctional TRAIL trimer facilitates cell-specific targeting and tumor cell killing

    doi: 10.1158/1535-7163.MCT-10-0225

    Figure Lengend Snippet: Human pancreatic cancer cells are killed in vivo following coinjection with TR3-coated RBCs

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, Maryland).

    Techniques: In Vivo

    Design and biochemical characterization of the genetically-encoded TRAIL trimer TR3

    Journal: Molecular cancer therapeutics

    Article Title: A genetically-encoded multifunctional TRAIL trimer facilitates cell-specific targeting and tumor cell killing

    doi: 10.1158/1535-7163.MCT-10-0225

    Figure Lengend Snippet: Design and biochemical characterization of the genetically-encoded TRAIL trimer TR3

    Article Snippet: To obtain concentrated TR3 protein stocks, the supernatants were applied to centrifugal filter devices with a 10 kDa molecular cut-off (Centricon Plus-20, Millipore, Bedford, Maryland).

    Techniques:

    SDS-PAGE ( A ) and immunoblot ( B ) analyses of recombinant Tb4CL4-like. (A): Lane 1, uninduced protein; lane 2, induced protein; lane 3, flow-through of protein sample; lane 4, elution of equilibration buffer; lanes 5–9, elution buffer from 20, 40, 100, 120 and 140 mM imidazole buffer, respectively. (B): Lane 1, Tb4CL4-like-His fusion protein was analyzed by immunoblotting with anti-His as the primary antibody. The target band is marked in red. The positions and sizes (in kDa) of the weight standards in lane M are indicated.

    Journal: Toxins

    Article Title: Cloning and Immunosuppressive Properties of an Acyl-Activating Enzyme from the Venom Apparatus of Tetrastichus brontispae (Hymenoptera: Eulophidae)

    doi: 10.3390/toxins11110672

    Figure Lengend Snippet: SDS-PAGE ( A ) and immunoblot ( B ) analyses of recombinant Tb4CL4-like. (A): Lane 1, uninduced protein; lane 2, induced protein; lane 3, flow-through of protein sample; lane 4, elution of equilibration buffer; lanes 5–9, elution buffer from 20, 40, 100, 120 and 140 mM imidazole buffer, respectively. (B): Lane 1, Tb4CL4-like-His fusion protein was analyzed by immunoblotting with anti-His as the primary antibody. The target band is marked in red. The positions and sizes (in kDa) of the weight standards in lane M are indicated.

    Article Snippet: SDS-PAGE and Immunoblot Analysis Protein concentration was determined with the Bradford assay kit (TransGen) and then concentrated to a proper range using ultrafiltration (Amicon Ultra 10 kDa, Millipore, Darmstadt, Germany) facilitated by centrifugation.

    Techniques: SDS Page, Recombinant, Flow Cytometry