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  • 91
    Thermo Fisher 37㠂 â
    Fcwf-4 Cornell University (CU) cells produce high titers of cell-free type I FIPV with rapid growth kinetics. A) Virus growth kinetics measured from cell-free supernatants of AK-D (black), Fcwf-4 ATCC (blue), and Fcwf-4 CU (red) cells infected with FIPV Black (MOI=0.1) over 96 hours. Arrows indicate time of peak titer presented as plaque-forming units (pfu)/mL. Titer determined by plaque assay on AK-D cells in triplicate; error bars ±SD. B) Cell-associated and cell-free virus titers were determined following infection of Fcwf-4 CU, AK-D, and Fcwf-4 ATCC cells with FIPV Black (MOI=0.1). Cell-free titer was determined from cell-clarified supernatants; cell-associated titer was determined from suspended cell monolayers following three freeze-thaw cycles alternating between −80°C and <t>37°C.</t> Samples were taken at hours post-infection (hpi) just prior to, at, and following the maximum (max) virus titer for each cell type. Titers determined by plaque assay on AK-D cells in triplicate; error bars ±SD.
    37㠂 â, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 124 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam 37㠂 â
    Fcwf-4 Cornell University (CU) cells produce high titers of cell-free type I FIPV with rapid growth kinetics. A) Virus growth kinetics measured from cell-free supernatants of AK-D (black), Fcwf-4 ATCC (blue), and Fcwf-4 CU (red) cells infected with FIPV Black (MOI=0.1) over 96 hours. Arrows indicate time of peak titer presented as plaque-forming units (pfu)/mL. Titer determined by plaque assay on AK-D cells in triplicate; error bars ±SD. B) Cell-associated and cell-free virus titers were determined following infection of Fcwf-4 CU, AK-D, and Fcwf-4 ATCC cells with FIPV Black (MOI=0.1). Cell-free titer was determined from cell-clarified supernatants; cell-associated titer was determined from suspended cell monolayers following three freeze-thaw cycles alternating between −80°C and <t>37°C.</t> Samples were taken at hours post-infection (hpi) just prior to, at, and following the maximum (max) virus titer for each cell type. Titers determined by plaque assay on AK-D cells in triplicate; error bars ±SD.
    37㠂 â, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Boehringer Mannheim 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Merck KGaA 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 91/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Sheldon Manufacturing 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Sheldon Manufacturing, used in various techniques. Bioz Stars score: 91/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Carl Zeiss 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Gemini Bio 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Gemini Bio, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Harvard Bioscience 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Harvard Bioscience, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Pinnacle Technology Inc 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Pinnacle Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Cellgro 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Cellgro, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Roche 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Clinical and Laboratory Standards Institute 37㠂 â
    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at <t>37°C</t> as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.
    37㠂 â, supplied by Clinical and Laboratory Standards Institute, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Pharmingen 37㠂 â
    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
    37㠂 â, supplied by Pharmingen, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Carl Roth GmbH 37㠂 â
    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
    37㠂 â, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Illumina Inc 37㠂 â
    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
    37㠂 â, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery 37㠂 â
    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
    37㠂 â, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck & Co 37㠂 â
    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
    37㠂 â, supplied by Merck & Co, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Corning Life Sciences 37㠂 â
    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
    37㠂 â, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 91/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco 37㠂 â
    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
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    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
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    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at <t>37°C</t> for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P
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    Image Search Results


    Fcwf-4 Cornell University (CU) cells produce high titers of cell-free type I FIPV with rapid growth kinetics. A) Virus growth kinetics measured from cell-free supernatants of AK-D (black), Fcwf-4 ATCC (blue), and Fcwf-4 CU (red) cells infected with FIPV Black (MOI=0.1) over 96 hours. Arrows indicate time of peak titer presented as plaque-forming units (pfu)/mL. Titer determined by plaque assay on AK-D cells in triplicate; error bars ±SD. B) Cell-associated and cell-free virus titers were determined following infection of Fcwf-4 CU, AK-D, and Fcwf-4 ATCC cells with FIPV Black (MOI=0.1). Cell-free titer was determined from cell-clarified supernatants; cell-associated titer was determined from suspended cell monolayers following three freeze-thaw cycles alternating between −80°C and 37°C. Samples were taken at hours post-infection (hpi) just prior to, at, and following the maximum (max) virus titer for each cell type. Titers determined by plaque assay on AK-D cells in triplicate; error bars ±SD.

    Journal: Virology

    Article Title: Characterizing replication kinetics and plaque production of type I feline infectious peritonitis virus in three feline cell lines

    doi: 10.1016/j.virol.2018.08.022

    Figure Lengend Snippet: Fcwf-4 Cornell University (CU) cells produce high titers of cell-free type I FIPV with rapid growth kinetics. A) Virus growth kinetics measured from cell-free supernatants of AK-D (black), Fcwf-4 ATCC (blue), and Fcwf-4 CU (red) cells infected with FIPV Black (MOI=0.1) over 96 hours. Arrows indicate time of peak titer presented as plaque-forming units (pfu)/mL. Titer determined by plaque assay on AK-D cells in triplicate; error bars ±SD. B) Cell-associated and cell-free virus titers were determined following infection of Fcwf-4 CU, AK-D, and Fcwf-4 ATCC cells with FIPV Black (MOI=0.1). Cell-free titer was determined from cell-clarified supernatants; cell-associated titer was determined from suspended cell monolayers following three freeze-thaw cycles alternating between −80°C and 37°C. Samples were taken at hours post-infection (hpi) just prior to, at, and following the maximum (max) virus titer for each cell type. Titers determined by plaque assay on AK-D cells in triplicate; error bars ±SD.

    Article Snippet: Cells were infected with 10-fold serial dilutions of viral samples for 1 h at 37°C, followed by overlaying with a 0.5% Oxoid agar (Oxoid LTD, #LP0028)-DMEM containing 1% FBS mixture.

    Techniques: Infection, Plaque Assay

    A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at 37°C as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Reconstitution of retroviral fusion and uncoating in a cell-free system

    doi: 10.1073/pnas.0401312101

    Figure Lengend Snippet: A cell-free system to study ASLV fusion and uncoating. ( A ) A schematic diagram of the cell-free system described in the text. ( B ) Low pH-dependent membrane fusion and added magnesium ions and dNTPs are critical for ASLV early DNA production. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 0.15 EGFP transducing unit. The ASLV fusion/uncoating reaction was then set up at 37°C as described in Materials and Methods with 150 μg of 293 S10 fraction and was performed under permissive (3 mM NH 4 Cl) or nonpermissive (63 mM NH 4 Cl) conditions for ASLV Env-dependent fusion. Samples were incubated with or without 3 mM MgCl 2 and 50 μM dNTPs. Viral DNA was then quantified by using a real-time PCR amplification method. The results are shown as the number of viral DNA molecules synthesized per cell that was used to make the VPNS. A representative example of an experiment that was performed at least three independent times, each time in triplicate, is shown, with the standard deviation of the data indicated with error bars.

    Article Snippet: All reactions were performed at 37°C for 6–12 h unless otherwise stated, and stopped by a 15-min incubation with 100 μg of Proteinase K (Boehringer Mannheim) at room temperature before shifting samples to -80°C.

    Techniques: Incubation, Real-time Polymerase Chain Reaction, Amplification, Synthesized, Standard Deviation

    A 293 cell nuclear extract promotes late ASLV DNA synthesis. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 1.0 EGFP transducing unit. Fusion/uncoating reactions were then set up at 37°C with 150 μg of 293 S10 fractions, 3 or 63 mM NH 4 Cl, and either with (+) or without (-) 1.6 × 10 6 cell equivalents of a 293 cell nuclear extract. Approximately 20 h later, early ( A ) and late ( B ) ASLV DNA products were measured by real-time PCR analysis as described in Materials and Methods . An experiment that was performed in triplicate is shown with the standard deviation of the data indicated with error bars. A similar ratio of early/late viral DNA molecules (600:1) was observed in a separate experiment that was performed in triplicate under the same conditions, except at a 20-fold lower apparent MOI (data not shown).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Reconstitution of retroviral fusion and uncoating in a cell-free system

    doi: 10.1073/pnas.0401312101

    Figure Lengend Snippet: A 293 cell nuclear extract promotes late ASLV DNA synthesis. 293(TVA800) cells were challenged with RCASBP(A)-EGFP at an estimated MOI of 1.0 EGFP transducing unit. Fusion/uncoating reactions were then set up at 37°C with 150 μg of 293 S10 fractions, 3 or 63 mM NH 4 Cl, and either with (+) or without (-) 1.6 × 10 6 cell equivalents of a 293 cell nuclear extract. Approximately 20 h later, early ( A ) and late ( B ) ASLV DNA products were measured by real-time PCR analysis as described in Materials and Methods . An experiment that was performed in triplicate is shown with the standard deviation of the data indicated with error bars. A similar ratio of early/late viral DNA molecules (600:1) was observed in a separate experiment that was performed in triplicate under the same conditions, except at a 20-fold lower apparent MOI (data not shown).

    Article Snippet: All reactions were performed at 37°C for 6–12 h unless otherwise stated, and stopped by a 15-min incubation with 100 μg of Proteinase K (Boehringer Mannheim) at room temperature before shifting samples to -80°C.

    Techniques: DNA Synthesis, Real-time Polymerase Chain Reaction, Standard Deviation

    NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at 37°C for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P

    Journal: The American Journal of Pathology

    Article Title: Neural Stem/Progenitor Cells Express Costimulatory Molecules That Are Differentially Regulated by Inflammatory and Apoptotic Stimuli

    doi:

    Figure Lengend Snippet: NSC can stimulate T proliferation in a B7-dependent fashion. A: Primary NSCs can elicit an allogeneic T cell response. NSC (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at 37°C for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic-purified T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. There was a significant increase in proliferation between NSC+T cells versus NSCs alone. ( P

    Article Snippet: For surface expression, proliferating NSCs were detached using incubation with EDTA for 30 minutes at 37°C, the cells were washed, counted and resuspended in staining buffer, treated with FC block (Pharmingen), and stained with labeled antibodies.

    Techniques: Incubation, Irradiation, Purification, Mouse Assay

    Stress-induced apoptosis differentially increases CD80 expression by NSCs and CD80 ligation increases apoptosis. A: Stress was induced by culturing NSCs with hydrogen peroxide at 100 μmol/L concentration for a 24 to 48 hour period ( left histograms ) or by media deprived of serum and growth factors (right histograms ). Cells were detached using EDTA-Versene, were incubated for 15 minutes at 37°C washed and counted for flow cytometry, 7-amino actinomycin D (7-ADD) staining was used to exclude non-viable cells. B: NSCs were incubated with TSA 100 ng/ml for 18 or 24 hours and examined by two-color staining with Annexin V and 7-AAD and anti-CD80 or anti-CD86 ( left dot blot ). The expression of B7 molecules was measured by gating on early apoptotic cells (Annexin-V positive, 7AAD negative) vs. live cells (Annexin and 7-AAD negative). The green background is the isotype control whereas the red shift is the anti-CD80 or anti-CD86 antibodies. The expression of CD80 is up-regulated in cells undergoing apoptosis ( left histograms ) as compared with CD86 where a slight up-regulation occurs after a 24-hour time period ( right histograms ). C: Actively proliferating C17.2 NSCs were left untreated, incubated with anti-CD80 (50 μg/ml), TSA (100 ng/ml), TSA plus anti-CD86, TSA plus anti-CD80 (5 to 10 μg/ml), TSA plus CD80 F(ab), TSA plus mIg, or TSA plus rat IgG2a and the percentage of apoptotic cells measured by Annexin and 7-AAD staining. Dot blots of apoptotic cell staining after 18 hours of culture comparing various culture conditions ( left ). The early apoptotic cells are seen in the lower right quadrant . D: Mean ± SE of percentage of apoptotic cells from triplicate cultures. Apoptotic cells were calculated as the sum of early and late apoptotic cells. These data were reproduced in three independent experiments (each done in triplicate). The effects of TSA alone is demonstrated by an increase in apoptotic cells compared to untreated cells ( P

    Journal: The American Journal of Pathology

    Article Title: Neural Stem/Progenitor Cells Express Costimulatory Molecules That Are Differentially Regulated by Inflammatory and Apoptotic Stimuli

    doi:

    Figure Lengend Snippet: Stress-induced apoptosis differentially increases CD80 expression by NSCs and CD80 ligation increases apoptosis. A: Stress was induced by culturing NSCs with hydrogen peroxide at 100 μmol/L concentration for a 24 to 48 hour period ( left histograms ) or by media deprived of serum and growth factors (right histograms ). Cells were detached using EDTA-Versene, were incubated for 15 minutes at 37°C washed and counted for flow cytometry, 7-amino actinomycin D (7-ADD) staining was used to exclude non-viable cells. B: NSCs were incubated with TSA 100 ng/ml for 18 or 24 hours and examined by two-color staining with Annexin V and 7-AAD and anti-CD80 or anti-CD86 ( left dot blot ). The expression of B7 molecules was measured by gating on early apoptotic cells (Annexin-V positive, 7AAD negative) vs. live cells (Annexin and 7-AAD negative). The green background is the isotype control whereas the red shift is the anti-CD80 or anti-CD86 antibodies. The expression of CD80 is up-regulated in cells undergoing apoptosis ( left histograms ) as compared with CD86 where a slight up-regulation occurs after a 24-hour time period ( right histograms ). C: Actively proliferating C17.2 NSCs were left untreated, incubated with anti-CD80 (50 μg/ml), TSA (100 ng/ml), TSA plus anti-CD86, TSA plus anti-CD80 (5 to 10 μg/ml), TSA plus CD80 F(ab), TSA plus mIg, or TSA plus rat IgG2a and the percentage of apoptotic cells measured by Annexin and 7-AAD staining. Dot blots of apoptotic cell staining after 18 hours of culture comparing various culture conditions ( left ). The early apoptotic cells are seen in the lower right quadrant . D: Mean ± SE of percentage of apoptotic cells from triplicate cultures. Apoptotic cells were calculated as the sum of early and late apoptotic cells. These data were reproduced in three independent experiments (each done in triplicate). The effects of TSA alone is demonstrated by an increase in apoptotic cells compared to untreated cells ( P

    Article Snippet: For surface expression, proliferating NSCs were detached using incubation with EDTA for 30 minutes at 37°C, the cells were washed, counted and resuspended in staining buffer, treated with FC block (Pharmingen), and stained with labeled antibodies.

    Techniques: Expressing, Ligation, Concentration Assay, Incubation, Flow Cytometry, Cytometry, Staining, Dot Blot

    NSC can stimulate CD8+ T cell proliferation and is dependent on CD80 and MHC class I expression. A: Primary NSCs cells (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at 37°C for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic purified CD8+ T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. Experiments were performed in quadruplicate. CTLA4Ig was used at a concentration of 5 μg/ml and anti-MHC-I was used at a concentration of 10 μg/ml. There was a significant increase in proliferation in NSC+ CD8+ T cells versus CD8+ T cells alone, ( P

    Journal: The American Journal of Pathology

    Article Title: Neural Stem/Progenitor Cells Express Costimulatory Molecules That Are Differentially Regulated by Inflammatory and Apoptotic Stimuli

    doi:

    Figure Lengend Snippet: NSC can stimulate CD8+ T cell proliferation and is dependent on CD80 and MHC class I expression. A: Primary NSCs cells (2 × 10 5 ) were stimulated with IFN-γ (100 U/ml) and incubated at 37°C for 24 hours, detached with Versene-EDTA then washed with HBSS 3% FBS and irradiated with 10000 R. Splenic purified CD8+ T cells (3 × 10 5 ) from SJL/J mice were used as responders. Cultures were pulsed with 3 H (1 μCi/well) after 72 hours and harvested at 96 hours. Experiments were performed in quadruplicate. CTLA4Ig was used at a concentration of 5 μg/ml and anti-MHC-I was used at a concentration of 10 μg/ml. There was a significant increase in proliferation in NSC+ CD8+ T cells versus CD8+ T cells alone, ( P

    Article Snippet: For surface expression, proliferating NSCs were detached using incubation with EDTA for 30 minutes at 37°C, the cells were washed, counted and resuspended in staining buffer, treated with FC block (Pharmingen), and stained with labeled antibodies.

    Techniques: Expressing, Incubation, Irradiation, Purification, Mouse Assay, Concentration Assay

    Primary neural stem cells are able to establish immunological synapses with T cells. A: Confocal phase microscopy shows a primary NSC-T cell conjugate and CD3 redistribution to the contact zone. NSC is stained green with nestin antibody and CD3 stained in red in a planar view. B: Three-dimensional reconstruction ( left ) and orthogonal view in the Z-plane ( right ) demonstrating the interaction between NSCs and T cells showing redistribution of CD3 to the contact zone ( arrows ). Quantitative profile analysis ( bottom ) by confocal microscopy shows intense expression of CD3 in the area of the contact zone (red peak ). Observe the intensity profile vector in white dotted line used for measurements of intensity in contact zone (white arrows ). C: Frequency of T cell-NSCs conjugates with or without pre-stimulation of NSCs with IFN-γ. NSCs were incubated with T cells for 20 minutes at 37°C and prepared for immunohistochemistry. The frequency of conjugates was measured by counting 100 NSCs in random fields per each well ( n = 3) in a volume of 1 ml.

    Journal: The American Journal of Pathology

    Article Title: Neural Stem/Progenitor Cells Express Costimulatory Molecules That Are Differentially Regulated by Inflammatory and Apoptotic Stimuli

    doi:

    Figure Lengend Snippet: Primary neural stem cells are able to establish immunological synapses with T cells. A: Confocal phase microscopy shows a primary NSC-T cell conjugate and CD3 redistribution to the contact zone. NSC is stained green with nestin antibody and CD3 stained in red in a planar view. B: Three-dimensional reconstruction ( left ) and orthogonal view in the Z-plane ( right ) demonstrating the interaction between NSCs and T cells showing redistribution of CD3 to the contact zone ( arrows ). Quantitative profile analysis ( bottom ) by confocal microscopy shows intense expression of CD3 in the area of the contact zone (red peak ). Observe the intensity profile vector in white dotted line used for measurements of intensity in contact zone (white arrows ). C: Frequency of T cell-NSCs conjugates with or without pre-stimulation of NSCs with IFN-γ. NSCs were incubated with T cells for 20 minutes at 37°C and prepared for immunohistochemistry. The frequency of conjugates was measured by counting 100 NSCs in random fields per each well ( n = 3) in a volume of 1 ml.

    Article Snippet: For surface expression, proliferating NSCs were detached using incubation with EDTA for 30 minutes at 37°C, the cells were washed, counted and resuspended in staining buffer, treated with FC block (Pharmingen), and stained with labeled antibodies.

    Techniques: Microscopy, Staining, Confocal Microscopy, Expressing, Plasmid Preparation, Incubation, Immunohistochemistry