cd70 cd27 interaction  (ATCC)


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    Structured Review

    ATCC cd70 cd27 interaction
    <t>CD70-specific</t> mAb l α hCD70 is a potent inhibitor of <t>CD27–CD70</t> interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells
    Cd70 Cd27 Interaction, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd70 cd27 interaction/product/ATCC
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd70 cd27 interaction - by Bioz Stars, 2024-04
    93/100 stars

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    1) Product Images from "CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants"

    Article Title: CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2013.555

    CD70-specific mAb l α hCD70 is a potent inhibitor of CD27–CD70 interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells
    Figure Legend Snippet: CD70-specific mAb l α hCD70 is a potent inhibitor of CD27–CD70 interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells

    Techniques Used: Incubation, Activity Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Derivative Assay, Binding Assay

    Initial characterization of scFv:CD70-TRAIL fusion proteins. ( a ) Scheme of scFv:l α hCD70-TRAIL fusion proteins. scFv:l α hCD70, CD70-specific scFv; F, Flag tag; TNC, tenascin-C trimerization domain; TRAIL, TRAILmutR1 and TRAILmutR2, aa 95–281 of wild-type TRAIL and TRAILR1- and TRAILR2-specific mutants derived thereof. ( b ) SDS-PAGE analysis of purified scFv:l α hCD70-TNC-TRAIL fusion proteins (100 ng each). ( c ) HT1080 cells were transiently transfected with a CD70 expression construct or empty vector. Next day, transfected cells were stimulated for an additional day with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2 in the presence and absence of a competing conventional CD70-specific antibody
    Figure Legend Snippet: Initial characterization of scFv:CD70-TRAIL fusion proteins. ( a ) Scheme of scFv:l α hCD70-TRAIL fusion proteins. scFv:l α hCD70, CD70-specific scFv; F, Flag tag; TNC, tenascin-C trimerization domain; TRAIL, TRAILmutR1 and TRAILmutR2, aa 95–281 of wild-type TRAIL and TRAILR1- and TRAILR2-specific mutants derived thereof. ( b ) SDS-PAGE analysis of purified scFv:l α hCD70-TNC-TRAIL fusion proteins (100 ng each). ( c ) HT1080 cells were transiently transfected with a CD70 expression construct or empty vector. Next day, transfected cells were stimulated for an additional day with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2 in the presence and absence of a competing conventional CD70-specific antibody

    Techniques Used: FLAG-tag, Derivative Assay, SDS Page, Purification, Transfection, Expressing, Construct, Plasmid Preparation

    CD70-restricted apoptosis induction by scFv:l α hCD70-TRAIL, scFv:l α hCD70-TRAILmutR1 and scFv:l α hCD70-TRAILmutR2. ( a ) The indicated cell lines were analyzed by using FACS for CD70 cell surface expression. ( b ) OVCAR3, Mino and Jurkat cells were cultured in 96-well plates, and half of the cells were pretreated with 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody. Cells were then challenged overnight in triplicates with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Finally, cellular viability was determined using the MTT assay or crystal violet staining. OVCAR3 cells were challenged in the presence of 2.5 μ g/ml CHX, which sensitizes this cell line for apoptosis induction. ( c ) The indicated cell line cells were stimulated in the presence and absence of the conventional CD70-specific antibody l α hCD70 for 4–6 h with 100 ng/ml of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Total cell lysates were analyzed by western blotting for processing of the indicated proteins
    Figure Legend Snippet: CD70-restricted apoptosis induction by scFv:l α hCD70-TRAIL, scFv:l α hCD70-TRAILmutR1 and scFv:l α hCD70-TRAILmutR2. ( a ) The indicated cell lines were analyzed by using FACS for CD70 cell surface expression. ( b ) OVCAR3, Mino and Jurkat cells were cultured in 96-well plates, and half of the cells were pretreated with 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody. Cells were then challenged overnight in triplicates with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Finally, cellular viability was determined using the MTT assay or crystal violet staining. OVCAR3 cells were challenged in the presence of 2.5 μ g/ml CHX, which sensitizes this cell line for apoptosis induction. ( c ) The indicated cell line cells were stimulated in the presence and absence of the conventional CD70-specific antibody l α hCD70 for 4–6 h with 100 ng/ml of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Total cell lysates were analyzed by western blotting for processing of the indicated proteins

    Techniques Used: Expressing, Cell Culture, MTT Assay, Staining, Western Blot

    Blockade of CD70 has no effect on cell death induction by conventional TRAIL. ( a ) Mino cells were cultured in 96-well plates, and half of the cells were pretreated with 5 μ g/ml of scFv:l α hCD70-TNC-GpL and then challenged overnight in triplicates with the indicated concentrations of TNC-TRAIL, TNC-TRAILmutR1 and TRAILmutR2 with and without anti-Flag (1 μ g/ml) oligomerization. Cellular viability was determined using the MTT assay. ( b ) FACS analysis of CD70 and CD27 expression of Raji and KMS12.BM cells. ( c ) Raji and KMS12.BM cells were cultured in 96-well plates in the presence and absence of 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody and were stimulated overnight in triplicates with the indicated concentrations of the various scFv:l α hCD70-TRAIL fusion proteins. Cellular viability was determined using the MTT assay
    Figure Legend Snippet: Blockade of CD70 has no effect on cell death induction by conventional TRAIL. ( a ) Mino cells were cultured in 96-well plates, and half of the cells were pretreated with 5 μ g/ml of scFv:l α hCD70-TNC-GpL and then challenged overnight in triplicates with the indicated concentrations of TNC-TRAIL, TNC-TRAILmutR1 and TRAILmutR2 with and without anti-Flag (1 μ g/ml) oligomerization. Cellular viability was determined using the MTT assay. ( b ) FACS analysis of CD70 and CD27 expression of Raji and KMS12.BM cells. ( c ) Raji and KMS12.BM cells were cultured in 96-well plates in the presence and absence of 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody and were stimulated overnight in triplicates with the indicated concentrations of the various scFv:l α hCD70-TRAIL fusion proteins. Cellular viability was determined using the MTT assay

    Techniques Used: Cell Culture, MTT Assay, Expressing

    cd70 cd27 interaction  (ATCC)


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    Structured Review

    ATCC cd70 cd27 interaction
    <t>CD70-specific</t> mAb l α hCD70 is a potent inhibitor of <t>CD27–CD70</t> interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells
    Cd70 Cd27 Interaction, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd70 cd27 interaction/product/ATCC
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd70 cd27 interaction - by Bioz Stars, 2024-04
    93/100 stars

    Images

    1) Product Images from "CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants"

    Article Title: CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2013.555

    CD70-specific mAb l α hCD70 is a potent inhibitor of CD27–CD70 interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells
    Figure Legend Snippet: CD70-specific mAb l α hCD70 is a potent inhibitor of CD27–CD70 interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells

    Techniques Used: Incubation, Activity Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Derivative Assay, Binding Assay

    Initial characterization of scFv:CD70-TRAIL fusion proteins. ( a ) Scheme of scFv:l α hCD70-TRAIL fusion proteins. scFv:l α hCD70, CD70-specific scFv; F, Flag tag; TNC, tenascin-C trimerization domain; TRAIL, TRAILmutR1 and TRAILmutR2, aa 95–281 of wild-type TRAIL and TRAILR1- and TRAILR2-specific mutants derived thereof. ( b ) SDS-PAGE analysis of purified scFv:l α hCD70-TNC-TRAIL fusion proteins (100 ng each). ( c ) HT1080 cells were transiently transfected with a CD70 expression construct or empty vector. Next day, transfected cells were stimulated for an additional day with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2 in the presence and absence of a competing conventional CD70-specific antibody
    Figure Legend Snippet: Initial characterization of scFv:CD70-TRAIL fusion proteins. ( a ) Scheme of scFv:l α hCD70-TRAIL fusion proteins. scFv:l α hCD70, CD70-specific scFv; F, Flag tag; TNC, tenascin-C trimerization domain; TRAIL, TRAILmutR1 and TRAILmutR2, aa 95–281 of wild-type TRAIL and TRAILR1- and TRAILR2-specific mutants derived thereof. ( b ) SDS-PAGE analysis of purified scFv:l α hCD70-TNC-TRAIL fusion proteins (100 ng each). ( c ) HT1080 cells were transiently transfected with a CD70 expression construct or empty vector. Next day, transfected cells were stimulated for an additional day with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2 in the presence and absence of a competing conventional CD70-specific antibody

    Techniques Used: FLAG-tag, Derivative Assay, SDS Page, Purification, Transfection, Expressing, Construct, Plasmid Preparation

    CD70-restricted apoptosis induction by scFv:l α hCD70-TRAIL, scFv:l α hCD70-TRAILmutR1 and scFv:l α hCD70-TRAILmutR2. ( a ) The indicated cell lines were analyzed by using FACS for CD70 cell surface expression. ( b ) OVCAR3, Mino and Jurkat cells were cultured in 96-well plates, and half of the cells were pretreated with 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody. Cells were then challenged overnight in triplicates with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Finally, cellular viability was determined using the MTT assay or crystal violet staining. OVCAR3 cells were challenged in the presence of 2.5 μ g/ml CHX, which sensitizes this cell line for apoptosis induction. ( c ) The indicated cell line cells were stimulated in the presence and absence of the conventional CD70-specific antibody l α hCD70 for 4–6 h with 100 ng/ml of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Total cell lysates were analyzed by western blotting for processing of the indicated proteins
    Figure Legend Snippet: CD70-restricted apoptosis induction by scFv:l α hCD70-TRAIL, scFv:l α hCD70-TRAILmutR1 and scFv:l α hCD70-TRAILmutR2. ( a ) The indicated cell lines were analyzed by using FACS for CD70 cell surface expression. ( b ) OVCAR3, Mino and Jurkat cells were cultured in 96-well plates, and half of the cells were pretreated with 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody. Cells were then challenged overnight in triplicates with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Finally, cellular viability was determined using the MTT assay or crystal violet staining. OVCAR3 cells were challenged in the presence of 2.5 μ g/ml CHX, which sensitizes this cell line for apoptosis induction. ( c ) The indicated cell line cells were stimulated in the presence and absence of the conventional CD70-specific antibody l α hCD70 for 4–6 h with 100 ng/ml of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Total cell lysates were analyzed by western blotting for processing of the indicated proteins

    Techniques Used: Expressing, Cell Culture, MTT Assay, Staining, Western Blot

    Blockade of CD70 has no effect on cell death induction by conventional TRAIL. ( a ) Mino cells were cultured in 96-well plates, and half of the cells were pretreated with 5 μ g/ml of scFv:l α hCD70-TNC-GpL and then challenged overnight in triplicates with the indicated concentrations of TNC-TRAIL, TNC-TRAILmutR1 and TRAILmutR2 with and without anti-Flag (1 μ g/ml) oligomerization. Cellular viability was determined using the MTT assay. ( b ) FACS analysis of CD70 and CD27 expression of Raji and KMS12.BM cells. ( c ) Raji and KMS12.BM cells were cultured in 96-well plates in the presence and absence of 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody and were stimulated overnight in triplicates with the indicated concentrations of the various scFv:l α hCD70-TRAIL fusion proteins. Cellular viability was determined using the MTT assay
    Figure Legend Snippet: Blockade of CD70 has no effect on cell death induction by conventional TRAIL. ( a ) Mino cells were cultured in 96-well plates, and half of the cells were pretreated with 5 μ g/ml of scFv:l α hCD70-TNC-GpL and then challenged overnight in triplicates with the indicated concentrations of TNC-TRAIL, TNC-TRAILmutR1 and TRAILmutR2 with and without anti-Flag (1 μ g/ml) oligomerization. Cellular viability was determined using the MTT assay. ( b ) FACS analysis of CD70 and CD27 expression of Raji and KMS12.BM cells. ( c ) Raji and KMS12.BM cells were cultured in 96-well plates in the presence and absence of 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody and were stimulated overnight in triplicates with the indicated concentrations of the various scFv:l α hCD70-TRAIL fusion proteins. Cellular viability was determined using the MTT assay

    Techniques Used: Cell Culture, MTT Assay, Expressing

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    ATCC cd70 cd27 interaction
    <t>CD70-specific</t> mAb l α hCD70 is a potent inhibitor of <t>CD27–CD70</t> interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells
    Cd70 Cd27 Interaction, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd70 cd27 interaction/product/ATCC
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd70 cd27 interaction - by Bioz Stars, 2024-04
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    CD70-specific mAb l α hCD70 is a potent inhibitor of CD27–CD70 interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells

    Journal: Cell Death & Disease

    Article Title: CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants

    doi: 10.1038/cddis.2013.555

    Figure Lengend Snippet: CD70-specific mAb l α hCD70 is a potent inhibitor of CD27–CD70 interaction. ( a ) OVCAR3 and Mino cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6. Cells were then incubated with 200 ng/ml CD27-Fc-GpL for an additional hour and after removal of unbound molecules cell-associated GpL activity was determined. ( b ) Mino and U266 cells were preincubated (1 h) with the indicated concentrations of the CD70-specific mAbs l α hCD70 and 1F6 and were then used to stimulate IL8 production in HT1080-CD27 cells that were cultured in a 96-well plate. Next day, cell culture supernatants were analyzed for the presence of IL8 by ELISA. ( c ) Monomeric and trimeric GpL fusion proteins of a l α hCD70-derived scFv were used for equilibrium binding studies with OVCAR3 and Mino cells

    Article Snippet: To obtain l α hCD70 (27B3), a Fab fragment phage library derived from llamas immunized with 786-O cells (ATCC) was screened for antibodies interfering with CD70-CD27 interaction and high affinity for CD70 essentially as described elsewhere.

    Techniques: Incubation, Activity Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Derivative Assay, Binding Assay

    Initial characterization of scFv:CD70-TRAIL fusion proteins. ( a ) Scheme of scFv:l α hCD70-TRAIL fusion proteins. scFv:l α hCD70, CD70-specific scFv; F, Flag tag; TNC, tenascin-C trimerization domain; TRAIL, TRAILmutR1 and TRAILmutR2, aa 95–281 of wild-type TRAIL and TRAILR1- and TRAILR2-specific mutants derived thereof. ( b ) SDS-PAGE analysis of purified scFv:l α hCD70-TNC-TRAIL fusion proteins (100 ng each). ( c ) HT1080 cells were transiently transfected with a CD70 expression construct or empty vector. Next day, transfected cells were stimulated for an additional day with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2 in the presence and absence of a competing conventional CD70-specific antibody

    Journal: Cell Death & Disease

    Article Title: CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants

    doi: 10.1038/cddis.2013.555

    Figure Lengend Snippet: Initial characterization of scFv:CD70-TRAIL fusion proteins. ( a ) Scheme of scFv:l α hCD70-TRAIL fusion proteins. scFv:l α hCD70, CD70-specific scFv; F, Flag tag; TNC, tenascin-C trimerization domain; TRAIL, TRAILmutR1 and TRAILmutR2, aa 95–281 of wild-type TRAIL and TRAILR1- and TRAILR2-specific mutants derived thereof. ( b ) SDS-PAGE analysis of purified scFv:l α hCD70-TNC-TRAIL fusion proteins (100 ng each). ( c ) HT1080 cells were transiently transfected with a CD70 expression construct or empty vector. Next day, transfected cells were stimulated for an additional day with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2 in the presence and absence of a competing conventional CD70-specific antibody

    Article Snippet: To obtain l α hCD70 (27B3), a Fab fragment phage library derived from llamas immunized with 786-O cells (ATCC) was screened for antibodies interfering with CD70-CD27 interaction and high affinity for CD70 essentially as described elsewhere.

    Techniques: FLAG-tag, Derivative Assay, SDS Page, Purification, Transfection, Expressing, Construct, Plasmid Preparation

    CD70-restricted apoptosis induction by scFv:l α hCD70-TRAIL, scFv:l α hCD70-TRAILmutR1 and scFv:l α hCD70-TRAILmutR2. ( a ) The indicated cell lines were analyzed by using FACS for CD70 cell surface expression. ( b ) OVCAR3, Mino and Jurkat cells were cultured in 96-well plates, and half of the cells were pretreated with 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody. Cells were then challenged overnight in triplicates with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Finally, cellular viability was determined using the MTT assay or crystal violet staining. OVCAR3 cells were challenged in the presence of 2.5 μ g/ml CHX, which sensitizes this cell line for apoptosis induction. ( c ) The indicated cell line cells were stimulated in the presence and absence of the conventional CD70-specific antibody l α hCD70 for 4–6 h with 100 ng/ml of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Total cell lysates were analyzed by western blotting for processing of the indicated proteins

    Journal: Cell Death & Disease

    Article Title: CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants

    doi: 10.1038/cddis.2013.555

    Figure Lengend Snippet: CD70-restricted apoptosis induction by scFv:l α hCD70-TRAIL, scFv:l α hCD70-TRAILmutR1 and scFv:l α hCD70-TRAILmutR2. ( a ) The indicated cell lines were analyzed by using FACS for CD70 cell surface expression. ( b ) OVCAR3, Mino and Jurkat cells were cultured in 96-well plates, and half of the cells were pretreated with 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody. Cells were then challenged overnight in triplicates with the indicated concentrations of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Finally, cellular viability was determined using the MTT assay or crystal violet staining. OVCAR3 cells were challenged in the presence of 2.5 μ g/ml CHX, which sensitizes this cell line for apoptosis induction. ( c ) The indicated cell line cells were stimulated in the presence and absence of the conventional CD70-specific antibody l α hCD70 for 4–6 h with 100 ng/ml of scFv:l α hCD70-TNC-TRAIL, scFv:l α hCD70-TNC-TRAILmutR1 and scFv:l α hCD70-TNC-TRAILmutR2. Total cell lysates were analyzed by western blotting for processing of the indicated proteins

    Article Snippet: To obtain l α hCD70 (27B3), a Fab fragment phage library derived from llamas immunized with 786-O cells (ATCC) was screened for antibodies interfering with CD70-CD27 interaction and high affinity for CD70 essentially as described elsewhere.

    Techniques: Expressing, Cell Culture, MTT Assay, Staining, Western Blot

    Blockade of CD70 has no effect on cell death induction by conventional TRAIL. ( a ) Mino cells were cultured in 96-well plates, and half of the cells were pretreated with 5 μ g/ml of scFv:l α hCD70-TNC-GpL and then challenged overnight in triplicates with the indicated concentrations of TNC-TRAIL, TNC-TRAILmutR1 and TRAILmutR2 with and without anti-Flag (1 μ g/ml) oligomerization. Cellular viability was determined using the MTT assay. ( b ) FACS analysis of CD70 and CD27 expression of Raji and KMS12.BM cells. ( c ) Raji and KMS12.BM cells were cultured in 96-well plates in the presence and absence of 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody and were stimulated overnight in triplicates with the indicated concentrations of the various scFv:l α hCD70-TRAIL fusion proteins. Cellular viability was determined using the MTT assay

    Journal: Cell Death & Disease

    Article Title: CD70-restricted specific activation of TRAILR1 or TRAILR2 using scFv-targeted TRAIL mutants

    doi: 10.1038/cddis.2013.555

    Figure Lengend Snippet: Blockade of CD70 has no effect on cell death induction by conventional TRAIL. ( a ) Mino cells were cultured in 96-well plates, and half of the cells were pretreated with 5 μ g/ml of scFv:l α hCD70-TNC-GpL and then challenged overnight in triplicates with the indicated concentrations of TNC-TRAIL, TNC-TRAILmutR1 and TRAILmutR2 with and without anti-Flag (1 μ g/ml) oligomerization. Cellular viability was determined using the MTT assay. ( b ) FACS analysis of CD70 and CD27 expression of Raji and KMS12.BM cells. ( c ) Raji and KMS12.BM cells were cultured in 96-well plates in the presence and absence of 10 μ g/ml of a scFv:l α hCD70 competing conventional CD70-specific antibody and were stimulated overnight in triplicates with the indicated concentrations of the various scFv:l α hCD70-TRAIL fusion proteins. Cellular viability was determined using the MTT assay

    Article Snippet: To obtain l α hCD70 (27B3), a Fab fragment phage library derived from llamas immunized with 786-O cells (ATCC) was screened for antibodies interfering with CD70-CD27 interaction and high affinity for CD70 essentially as described elsewhere.

    Techniques: Cell Culture, MTT Assay, Expressing